KLF4 Antibody

What is KLF4 and why is it important in biological research?

KLF4 (Kruppel-like factor 4, also known as GKLF or EZF) is a zinc finger transcription factor that activates or represses gene expression through interaction with specific co-activators such as the p300-CBP coactivator family. In embryonic stem cells, KLF4 serves as a critical indicator of stemness capacity and is one of the four Yamanaka factors used in cellular reprogramming to generate induced pluripotent stem cells. KLF4 is also involved in cell cycle regulation, differentiation, and apoptosis across various tissue types. The importance of KLF4 in biological research stems from its roles in embryonic development, tissue homeostasis, cancer progression, and stem cell biology, making KLF4 antibodies valuable tools for investigating these processes .

What applications are KLF4 antibodies commonly used for?

KLF4 antibodies have been validated for multiple research applications:

How do I select the appropriate KLF4 antibody for my experiment?

Selection of a KLF4 antibody should be based on several criteria specific to your experimental needs:

• Species reactivity: Ensure the antibody recognizes KLF4 in your species of interest (human, mouse, rat, etc.). Cross-reactivity information is typically provided in product documentation.

• Application validation: Verify that the antibody has been validated for your specific application (WB, IHC, IF, ChIP, etc.) with published validation data.

• Clonality: Polyclonal antibodies often provide higher sensitivity but potentially lower specificity, while monoclonal antibodies offer greater specificity for a single epitope.

• Immunogen information: Check whether the antibody was raised against a synthetic peptide, recombinant protein, or fusion protein, as this can affect recognition of native versus denatured protein.

• Antibody host species: Consider potential cross-reactivity issues with secondary antibodies in your experimental system.
  Many commercial KLF4 antibodies have been validated in multiple applications and species, but verification through published literature and documented validation data is essential for experimental success .

What are the optimal conditions for Western blot detection of KLF4?

For optimal Western blot detection of KLF4:

• Sample preparation: Use RIPA or similar lysis buffers containing protease inhibitors. KLF4 can be detected in various cell lines, including HT-29, SW480, HCT-116, A431, A549, and HUVEC cells, as well as mouse liver and lung tissues.

• Gel conditions: Use 10-12% SDS-PAGE gels under reducing conditions.

• Protein amount: Load 20-50 μg of total protein per lane.

• Transfer conditions: Transfer to PVDF membrane (recommended over nitrocellulose for KLF4).

• Blocking: 5% non-fat dry milk or BSA in TBST for 1 hour at room temperature.

• Primary antibody: Dilute KLF4 antibody to 0.5-1 μg/mL (approximately 1:1000-1:8000 dilution) in blocking buffer and incubate overnight at 4°C.

• Secondary antibody: Use HRP-conjugated secondary antibody (specific to the host species of the primary antibody) at 1:2000-1:10000 dilution.

• Detection: ECL substrate systems work well for KLF4 detection.

• Expected band size: KLF4 typically appears at approximately 50-60 kDa, though the calculated molecular weight is 54 kDa.
  Using Immunoblot Buffer Group 5 has shown good results for KLF4 detection in validated protocols .

How should I perform immunohistochemistry for KLF4 in tissue samples?

For IHC detection of KLF4 in tissue samples:

• Tissue preparation: Use formalin-fixed, paraffin-embedded (FFPE) sections (4-6 μm thickness).

• Antigen retrieval: Perform heat-induced epitope retrieval using TE buffer at pH 9.0 (preferred) or citrate buffer at pH 6.0 as an alternative.

• Blocking: 5-10% normal serum (from the same species as the secondary antibody) in PBS for 1 hour at room temperature.

• Primary antibody: Apply KLF4 antibody at 15 μg/mL (approximately 1:50-1:500 dilution) and incubate overnight at 4°C.

• Detection system: Use HRP-DAB Cell & Tissue Staining Kit or equivalent detection system.

• Counterstaining: Hematoxylin provides good nuclear counterstaining for KLF4 IHC.

• Expected localization: KLF4 is typically localized to the nuclei of epithelial cells, especially in colon tissue.

• Controls: Include appropriate positive controls (human colon, tonsil, or spleen tissues) and negative controls (primary antibody omission or isotype control).
  This protocol has been validated for human and mouse tissues, with specific labeling observed in the nuclei of epithelial cells in human colon samples .

What is the best protocol for ChIP experiments using KLF4 antibodies?

For ChIP experiments targeting KLF4:

• Cell preparation: Harvest 5-10 x 10^6 cells per ChIP reaction (embryonic stem cells like BG01V human embryonic stem cells work well).

• Crosslinking: Fix cells with 1% formaldehyde for 10 minutes at room temperature, followed by quenching with 125 mM glycine.

• Cell lysis: Resuspend cells in appropriate lysis buffer with protease inhibitors.

• Chromatin shearing: Sonicate to generate DNA fragments of 200-1000 bp (optimization required for each sonicator).

• Immunoprecipitation:

  • Use 5 μg of KLF4 antibody per reaction

  • Include appropriate control antibody (e.g., IgG from same species as KLF4 antibody)

  • Incubate overnight at 4°C with rotation

• Collection of immunocomplexes: Use biotinylated secondary antibody (e.g., Biotinylated Anti-Goat IgG) followed by streptavidin ferrofluid (e.g., MagCellect Streptavidin Ferrofluid).

• Washing: Perform stringent washes to remove non-specific binding.

• DNA purification: Elute and purify DNA using chelating resin solution or commercial kits.

• Analysis: Perform qPCR or standard PCR with primers specific to your genomic regions of interest. The B2R promoter has been successfully detected in KLF4 ChIP experiments.
  This protocol has been validated for detecting KLF4-regulated genes in embryonic stem cells, providing insights into transcriptional regulation mechanisms .

How can I use KLF4 antibodies to study cellular reprogramming and stemness?

KLF4 antibodies provide valuable tools for investigating cellular reprogramming and stemness through several advanced applications:

• Tracking reprogramming dynamics: Monitor KLF4 expression levels and subcellular localization during iPSC generation using immunofluorescence at different time points.

• Co-immunoprecipitation (Co-IP): Identify KLF4 interaction partners during reprogramming by performing Co-IP with KLF4 antibodies followed by mass spectrometry analysis.

• ChIP-seq analysis: Combine ChIP using KLF4 antibodies with next-generation sequencing to map genome-wide KLF4 binding sites in stem cells versus differentiated cells.

• Sequential ChIP (ChIP-reChIP): Investigate co-occupancy of KLF4 with other pluripotency factors (OCT4, SOX2, NANOG) at specific genomic loci.

• Immunofluorescence co-localization: Perform dual immunostaining of KLF4 with other stemness markers in embryonic stem cells using appropriate KLF4 antibodies (10 μg/mL concentration has been validated for BG01V human embryonic stem cells).

• Multi-parameter flow cytometry: Combine surface stemness markers with intracellular KLF4 staining to characterize stem cell populations at single-cell resolution.
  These approaches enable researchers to dissect the molecular mechanisms through which KLF4 contributes to pluripotency maintenance and cellular reprogramming, with validated protocols available using specific antibodies like those described in the search results .

What strategies can resolve conflicting KLF4 antibody data between different experimental systems?

Resolving conflicting KLF4 antibody data requires systematic troubleshooting and validation:

• Epitope mapping: Different antibodies may recognize distinct epitopes that could be differentially affected by post-translational modifications or protein-protein interactions. Compare immunogen sequences between antibodies showing discrepant results.

• Isoform specificity: KLF4 can exist in multiple isoforms. Verify whether your antibody recognizes all isoforms or is specific to particular variants by examining the immunogen sequence and comparing with known isoform structures.

• Antibody validation using genetic approaches:

  • Perform siRNA/shRNA knockdown of KLF4 to confirm specificity

  • Use CRISPR/Cas9-mediated KLF4 knockout cells as negative controls

  • Overexpress tagged KLF4 and verify co-detection with both anti-tag and anti-KLF4 antibodies

• Cross-platform validation: If results are inconsistent between applications (e.g., WB vs. IHC), validate findings using orthogonal techniques such as qPCR, ELISA, or mass spectrometry.

• Technical parameter adjustment: Modify fixation conditions, antigen retrieval methods, or antibody concentrations based on cell/tissue type. For example, some antibodies work better with TE buffer pH 9.0 for antigen retrieval, while others may require citrate buffer pH 6.0.

• Positive and negative control samples: Include cell lines with known KLF4 expression levels (positive: HT-29, SW480, HCT-116; negative: HDLM-2) across experiments to normalize and calibrate results.
  Studies have shown that different cellular contexts can affect antibody performance, making systematic validation crucial for accurate KLF4 detection and quantification .

How can I quantitatively assess KLF4 protein levels in complex tissue samples?

For quantitative assessment of KLF4 protein levels in complex tissues:

• Simple Western automated capillary immunoassay: This technique provides high-sensitivity, quantitative protein detection with minimal sample requirements. For KLF4, use 25 μg/mL antibody concentration against 0.2 mg/mL protein lysate for reliable detection at approximately 63 kDa.

• Quantitative immunofluorescence:

  • Use tile scanning confocal microscopy with consistent laser and detector settings

  • Include calibration standards in each experiment

  • Apply automated image analysis with nuclear segmentation

  • Normalize KLF4 signal intensity to nuclear area or DAPI intensity

  • Use software packages like CellProfiler or QuPath for quantification

• Multiplex immunohistochemistry: Combine KLF4 antibody with markers for specific cell types to quantify KLF4 expression in distinct cell populations within heterogeneous tissues.

• Tissue microarray analysis: Apply KLF4 antibodies (15 μg/mL concentration) to tissue microarrays containing multiple patient samples to compare expression levels across different pathological conditions.

• Laser capture microdissection combined with Western blotting: Isolate specific regions from complex tissues before protein extraction and quantitative Western blot analysis.

• Mass spectrometry-based validation: Use targeted mass spectrometry approaches like selected reaction monitoring (SRM) to validate and quantify KLF4 protein levels independently of antibody-based methods.
  These approaches enable precise quantification of KLF4 expression in specific cell populations within complex tissues, facilitating more nuanced understanding of its biological roles in development and disease .

How can I resolve non-specific banding patterns in KLF4 Western blots?

Non-specific banding in KLF4 Western blots can be addressed through several optimization strategies:

• Antibody concentration adjustment: Titrate KLF4 antibody concentration to find the optimal balance between specific signal and background. Start with the recommended 0.5 μg/mL and adjust as needed.

• Blocking optimization: Test different blocking agents (BSA, non-fat dry milk, commercial blockers) and concentrations (3-5%) to reduce non-specific binding.

• Buffer composition: Use Immunoblot Buffer Group 5 as recommended for KLF4 antibodies, which has been validated to produce specific bands at approximately 60 kDa.

• Sample preparation: Ensure complete denaturation of proteins by boiling samples in reducing sample buffer for 5-10 minutes before loading.

• Washing stringency: Increase the number and duration of washing steps with TBST to remove unbound antibody.

• Controls for specificity:

  • Include lysates from cell lines with known KLF4 expression (positive: HT-29, SW480, HCT-116)

  • Include lysates from KLF4-negative cells as negative controls (e.g., HDLM-2)

  • Perform antibody pre-absorption with immunizing peptide if available

• Secondary antibody optimization: Use highly cross-adsorbed secondary antibodies to minimize cross-reactivity with endogenous immunoglobulins.

• Expected molecular weight: KLF4 typically appears at 50-60 kDa despite a calculated molecular weight of 54 kDa. Post-translational modifications may cause slight variation in apparent molecular weight.
  These optimizations have proven effective in producing clean, specific KLF4 detection in Western blot applications across multiple cell types .

What are the key considerations for optimizing KLF4 immunofluorescence staining?

For optimal KLF4 immunofluorescence staining:

• Fixation method optimization:

  • Test different fixation reagents (4% paraformaldehyde, methanol, acetone)

  • Optimize fixation duration (10-30 minutes)

  • For KLF4, immersion fixation has been validated for successful staining

• Antigen retrieval:

  • For tissue sections or stubborn epitopes, heat-induced epitope retrieval may be necessary

  • Compare citrate buffer (pH 6.0) versus TE buffer (pH 9.0)

• Permeabilization conditions:

  • Optimize detergent type and concentration (0.1-0.5% Triton X-100 or 0.1-0.3% Saponin)

  • Adjust permeabilization time based on cell type and fixation method

• Antibody concentration and incubation:

  • Start with 5-15 μg/mL KLF4 antibody as recommended

  • Test both short incubations at room temperature (3 hours) and overnight incubation at 4°C

  • For stem cells, 10 μg/mL for 3 hours at room temperature has been validated

• Signal amplification options:

  • Use fluorophore-conjugated secondary antibodies with appropriate brightness

  • NorthernLights 557-conjugated Anti-Goat IgG has been validated for KLF4 detection

  • Consider tyramide signal amplification for low-abundance KLF4 detection

• Counterstaining:

  • DAPI for nuclear visualization works well with KLF4 staining

  • Consider additional markers for co-localization studies

• Mounting media selection:

  • Use anti-fade mounting media to prevent photobleaching

  • Select mounting media compatible with your fluorophore

• Expected localization pattern:

  • KLF4 typically shows nuclear localization in stem cells and epithelial cells

  • Some cytoplasmic staining may be observed in certain cell types
    Following these guidelines has produced specific KLF4 staining in various cell types, including embryonic stem cells and colon cancer cell lines .

What controls are essential for validating KLF4 antibody specificity?

Essential controls for validating KLF4 antibody specificity include:

• Genetic controls:

  • siRNA/shRNA knockdown of KLF4: Should show corresponding reduction in signal intensity

  • CRISPR/Cas9 KLF4 knockout cells: Should show absence of specific signal

  • KLF4 overexpression: Should show increased signal intensity proportional to expression level

• Peptide competition assay:

  • Pre-incubate KLF4 antibody with immunizing peptide/protein

  • Should abolish or significantly reduce specific signal

• Cell type controls:

  • Positive control cells with known KLF4 expression:

    • HT-29, SW480, HCT-116 human colorectal cell lines

    • BG01V human embryonic stem cells

  • Negative control cells with minimal KLF4 expression:

    • HDLM-2 Human Hodgkin's Lymphoma Cell Line has been validated as a negative control

• Tissue controls:

  • Human colon tissue: Shows specific nuclear labeling in epithelial cells

  • Human tonsil and spleen: Also show KLF4 expression

  • Include these as positive controls in IHC experiments

• Technical controls:

  • Isotype control antibody: Same species and isotype as KLF4 antibody

  • Secondary antibody only: To assess non-specific binding of secondary antibody

  • Omission of primary antibody: To assess autofluorescence or non-specific binding

• Orthogonal validation:

  • Compare results with multiple KLF4 antibodies recognizing different epitopes

  • Validate protein expression using independent methods (RT-qPCR, mass spectrometry)
    These validation controls are essential for confirming the specificity of KLF4 antibody staining and establishing confidence in experimental results, particularly when investigating complex biological systems .

How can I utilize KLF4 antibodies to investigate epigenetic regulation mechanisms?

KLF4 antibodies can be powerful tools for investigating epigenetic regulation through several specialized approaches:

• ChIP-seq for genome-wide binding profiles:

  • Use 5 μg KLF4 antibody per 5 x 10^6 cells as validated in protocols

  • Map KLF4 binding sites across the genome in different cell states

  • Integrate with histone modification and DNA methylation data to understand epigenetic context of KLF4 binding

• Sequential ChIP (ChIP-reChIP):

  • Perform first ChIP with KLF4 antibody

  • Elute complexes and perform second ChIP with antibodies against chromatin modifiers

  • Identify genomic regions where KLF4 co-occupies with specific epigenetic regulators

• ChIP-MS (Chromatin Immunoprecipitation coupled with Mass Spectrometry):

  • Use KLF4 antibodies to pull down chromatin complexes

  • Identify protein interactors through mass spectrometry

  • Map KLF4 interactions with epigenetic machinery

• Proximity ligation assay (PLA):

  • Detect in situ protein-protein interactions between KLF4 and epigenetic regulators

  • Visualize and quantify molecular proximities at single-molecule resolution

• CUT&RUN or CUT&Tag with KLF4 antibodies:

  • Higher signal-to-noise alternative to ChIP

  • Requires less starting material than traditional ChIP

  • Can be adapted using validated KLF4 antibodies

• Dual immunofluorescence with epigenetic marks:

  • Co-stain for KLF4 (using 5-15 μg/mL antibody) and histone modifications

  • Analyze correlation between KLF4 expression and epigenetic states at single-cell level
    Research has demonstrated that KLF4 interacts with histone deacetylases (HDACs), specifically showing that KLF4 can rescue effects of HDAC1 on leukemia cell proliferation, suggesting important epigenetic regulatory mechanisms. These techniques enable detailed investigation of how KLF4 influences and is influenced by the epigenetic landscape .

What are the best approaches for studying KLF4 in cancer development and progression?

For studying KLF4 in cancer development and progression:

• Tissue microarray analysis:

  • Apply KLF4 antibodies (15 μg/mL) to tumor tissue microarrays

  • Correlate KLF4 expression patterns with clinicopathological parameters

  • Human colon tissue shows specific nuclear KLF4 labeling in epithelial cells, providing a baseline for comparison with malignant transformation

• Cancer cell line profiling:

  • Compare KLF4 expression across cancer cell lines using Western blot (0.5 μg/mL antibody)

  • Validated cell lines include HT-29, SW480, and HCT-116 colorectal cancer lines

  • Correlate expression with proliferation, invasion, and drug resistance phenotypes

• Functional studies with antibody validation:

  • Perform KLF4 knockdown/overexpression in cancer cells

  • Validate knockdown efficiency using validated KLF4 antibodies

  • Assess effects on cell cycle regulators (p21, p27) that have been shown to be regulated by KLF4

• Chromatin immunoprecipitation for cancer-specific targets:

  • Identify KLF4 target genes in cancer cells using ChIP with 5 μg antibody per 5 x 10^6 cells

  • Compare binding profiles between normal and cancer cells

  • Validate binding to promoters of key cancer-related genes (e.g., p21, p27)

• Single-cell analysis of tumor heterogeneity:

  • Use flow cytometry with appropriate KLF4 antibodies (0.40 μg per 10^6 cells)

  • Combine with cancer stem cell markers

  • Sort subpopulations for functional characterization

• Circulating tumor cell detection:

  • Adapt immunocytochemistry protocols (5-15 μg/mL antibody concentration)

  • Combine with epithelial markers for CTC identification

• Patient-derived xenograft (PDX) models:

  • Apply validated IHC protocols (15 μg/mL antibody) to PDX tissue sections

  • Monitor KLF4 expression during tumor progression and treatment response
    These approaches can reveal the context-dependent roles of KLF4 in cancer, as it can function as both an oncogene and tumor suppressor depending on tissue type and genetic context .

How can I adapt KLF4 antibody protocols for high-throughput or automated platforms?

Adapting KLF4 antibody protocols for high-throughput or automated platforms requires several key modifications:

• Automated Western blot systems:

  • Simple Western capillary-based immunoassay:

    • Use 25 μg/mL KLF4 antibody concentration as validated

    • Expect KLF4 detection at approximately 63 kDa

    • Compatible with 12-230 kDa separation systems

    • Requires minimal sample volume (0.2 mg/mL protein concentration)

• High-content imaging systems:

  • Optimization for 96/384-well formats:

    • Reduce primary KLF4 antibody concentration to 2-5 μg/mL

    • Shorten incubation times (1-2 hours at room temperature)

    • Use fluorophore-conjugated secondary antibodies optimized for automated imaging

  • Multiplexing capabilities:

    • Combine KLF4 antibody with cell cycle markers

    • Use spectrally distinct fluorophores for co-detection

• Automated immunohistochemistry platforms:

  • Protocol adaptation:

    • Standardize antigen retrieval (TE buffer pH 9.0)

    • Use 5-10 μg/mL KLF4 antibody concentration

    • Optimize detection chemistry for platform-specific requirements

  • Quality control measures:

    • Include positive control tissues (human colon) on each run

    • Implement automated scoring algorithms

• Microfluidic ChIP systems:

  • Scale down protocol:

    • Reduce cell input to 1 x 10^6 cells

    • Maintain antibody:chromatin ratio (1 μg antibody per 10^6 cells)

    • Optimize washing steps for microfluidic format

• High-throughput flow cytometry:

  • Protocol optimization:

    • Standard 0.40 μg KLF4 antibody per 10^6 cells

    • Reduce incubation time to 30-60 minutes

    • Implement automated compensation and gating strategies

• Automated liquid handling integration:

  • Buffer standardization:

    • Use commercial buffer systems compatible with liquid handlers

    • Implement positive displacement pipetting for viscous solutions

  • Protocol segmentation:

    • Break protocols into discrete steps optimized for liquid handling

    • Include adequate mixing steps between reagent additions
      These adaptations facilitate large-scale studies of KLF4 biology across multiple samples or conditions while maintaining the specificity and sensitivity established in manual protocols .

What emerging technologies are advancing KLF4 antibody applications in research?

Several emerging technologies are expanding the capabilities of KLF4 antibody applications in cutting-edge research:

• CUT&RUN and CUT&Tag:

  • Adaptable with validated KLF4 antibodies

  • Offers higher resolution mapping of KLF4 genomic binding sites

  • Requires significantly less starting material than traditional ChIP

  • Provides cleaner signal-to-noise ratios for more precise binding site identification

• Spatial transcriptomics combined with KLF4 immunodetection:

  • Correlates KLF4 protein expression with transcriptional landscapes

  • Preserves tissue architecture and cellular context

  • Reveals spatial relationships between KLF4-expressing cells and their microenvironment

• Single-cell proteomics with antibody-based detection:

  • Mass cytometry (CyTOF) with metal-conjugated KLF4 antibodies

  • Single-cell Western blotting for KLF4 quantification

  • Microfluidic antibody capture for single-cell protein analysis

• Live-cell imaging with KLF4 nanobodies:

  • Smaller antibody derivatives for reduced interference with protein function

  • Compatible with fluorescent protein fusions for dynamic tracking

  • Enables real-time monitoring of KLF4 dynamics during cellular processes

• CRISPR-based tagging combined with antibody detection:

  • Endogenous tagging of KLF4 for improved antibody specificity

  • Eliminates artifacts associated with overexpression systems

  • Maintains native regulatory mechanisms

• AI-enhanced image analysis for KLF4 quantification:

  • Deep learning algorithms for automated quantification

  • Improved sensitivity for detecting subtle expression changes

  • More objective analysis across large datasets

• Proximity labeling coupled with KLF4 antibody validation:

  • BioID or APEX2 fusions with KLF4

  • Maps protein-protein interactions in native contexts

  • Validated using established KLF4 antibodies
    These technologies expand the toolbox available for KLF4 research, enabling more nuanced understanding of its functions across different biological contexts and disease states .

What are the most critical factors for successful reproducibility when using KLF4 antibodies?

Ensuring reproducibility with KLF4 antibodies requires attention to several critical factors:

• Antibody validation documentation:

  • Select antibodies with comprehensive validation data across multiple applications

  • Review knockout/knockdown validation data where available

  • Verify species cross-reactivity claims with experimental evidence

• Lot-to-lot consistency:

  • Test new antibody lots against previous lots when possible

  • Maintain reference samples for comparative analysis

  • Document lot numbers in protocols and publications

• Standardized protocols with detailed parameters:

  • Document exact antibody concentrations (e.g., 0.5 μg/mL for WB, 15 μg/mL for IHC)

  • Specify buffer compositions and pH values

  • Record incubation times and temperatures precisely

  • Note that KLF4 detection works optimally with specific conditions (e.g., TE buffer pH 9.0 for antigen retrieval)

• Comprehensive controls:

  • Include biological positive controls (HT-29, SW480, HCT-116 cells)

  • Include biological negative controls (HDLM-2 cells)

  • Implement technical controls (isotype, secondary-only)

• Cell/tissue preparation consistency:

  • Standardize fixation protocols (duration, temperature, reagent source)

  • Control cell culture conditions (passage number, confluence)

  • Normalize protein loading amounts

• Imaging and detection standardization:

  • Maintain consistent instrument settings between experiments

  • Include calibration standards for quantitative analyses

  • Document image acquisition parameters

• Data analysis transparency:

  • Report raw data alongside processed results

  • Document analysis pipelines and parameters

  • Share analysis code when possible
    Adhering to these principles ensures that findings related to KLF4 expression, localization, and function can be reliably reproduced across different laboratories and experimental settings, advancing our collective understanding of this important transcription factor .

How can researchers integrate KLF4 antibody data with other omics approaches for comprehensive biological insights?

Integrating KLF4 antibody data with other omics approaches provides multi-dimensional insights into biological processes:

• ChIP-seq and RNA-seq integration:

  • Map KLF4 binding sites using ChIP-seq with validated antibodies (5 μg per 5 x 10^6 cells)

  • Correlate binding sites with transcriptional changes from RNA-seq

  • Identify direct KLF4 targets versus secondary effects

  • The B2R promoter has been validated as a KLF4 target through ChIP analysis

• Proteomics and KLF4 antibody-based data:

  • Compare KLF4 protein levels detected by antibodies with global proteome changes

  • Identify post-translational modifications affecting KLF4 function

  • Discover protein interaction networks through IP-MS using validated KLF4 antibodies

• Single-cell multi-omics:

  • Combine KLF4 protein detection (using flow cytometry with 0.40 μg antibody per 10^6 cells)

  • Integrate with scRNA-seq from the same cell populations

  • Correlate protein and mRNA expression at single-cell resolution

• Spatial biology integration:

  • Map KLF4 expression in tissue contexts using IHC/IF (15 μg/mL antibody concentration)

  • Overlay with spatial transcriptomics data

  • Correlate with tissue microenvironment features

• Functional genomics correlation:

  • Validate CRISPR/RNAi screens with KLF4 antibody-based expression analysis

  • Connect genetic dependencies with KLF4 regulatory networks

  • Studies have shown KLF4's role in regulating cell cycle genes like p21 and p27

• Epigenomic integration:

  • Correlate KLF4 binding (from ChIP) with histone modifications and chromatin accessibility

  • Develop predictive models of KLF4 function based on chromatin context

  • KLF4 has been shown to interact with histone modifiers like HDAC1

• Clinical sample correlation:

  • Analyze KLF4 expression in patient samples using validated antibody protocols

  • Correlate with clinical parameters and outcomes

  • Integrate with patient genomic and transcriptomic data
    This multi-omics integration approach provides a comprehensive understanding of KLF4 biology across different contexts, from molecular mechanisms to physiological functions and disease relevance, creating a more complete picture than any single approach alone .