KLF4 Antibody
Description
Cancer Biology
-
Tumor Suppression: KLF4 overexpression in nasopharyngeal carcinoma (NPC) cells reduces tumor growth by upregulating p21 (a cell cycle inhibitor) and lowering proliferation markers (BrdU, Ki67) .
-
Metastasis Promotion: Knockdown of KLF4 in HONE1 NPC cells increases lung metastasis in mice by 5-fold compared to controls .
| Parameter | KLF4-Expressing Cells | KLF4-Depleted Cells |
|---|---|---|
| Tumor volume (mm³) | 150 ± 20 | 320 ± 30 |
| Lung metastasis nodules | 6.3 ± 3.3 | 31.4 ± 12.7 |
| p21 expression | ↑ 3.5-fold | ↓ 2.8-fold |
Immune Regulation
-
KLF4 mediates efferocytosis (clearance of apoptotic cells) in macrophages, enhancing injury resolution via trained immunity .
-
It suppresses NF-κB-driven inflammation in endothelial cells but promotes it in macrophages, highlighting context-dependent roles .
Stem Cell Research
-
KLF4 is a marker of pluripotency in embryonic stem cells (ESCs) and aids reprogramming somatic cells into induced pluripotent stem cells (iPSCs) .
-
In chromatin immunoprecipitation (ChIP), AF3640 confirmed KLF4 binding to the B2R promoter in BG01V ESCs .
Validation and Quality Control
-
Specificity: AF3640 shows no cross-reactivity in HDLM-2 lymphoma cells (KLF4-negative) .
-
Functional Assays: Knockdown/rescue experiments in leukemia cells demonstrated KLF4’s role in antagonizing HDAC1-induced proliferation .
Emerging Applications
Product Specs
Target Background
- Downstream targets of KLF4-mCpG binding, such as guanine nucleotide exchange factors, act as effectors of KLF4-induced mitochondrial fusion, cell cycle arrest, and cell protection. PMID: 29507094
- Human and mouse WNT10A mutant palmoplantar and tongue epithelia exhibit specific differentiation defects that are mirrored by the loss of the transcription factor KLF4. PMID: 28589954
- KLF4 might serve as a potential marker to predict prognosis in solid cancer patients. PMID: 29940144
- Overexpression of SOX5 increases Kruppel-like factor 4 (KLF4) gene expression, while silencing SOX5 decreases it. Knockdown of KLF4 abolishes the suppressive effect of SOX5 overexpression on osteogenic differentiation of human mesenchymal stem cells (hMSCs). PMID: 29890823
- SIRT1 interacts with and deacetylates KLF4. Deacetylation by SIRT1 promotes nuclear accumulation of KLF4 and enhances its binding to the CLDN5 promoter in the nucleus. PMID: 28888043
- miR-145 promotes hepatic stellate cell activation and liver fibrosis by targeting KLF4. PMID: 28091538
- These findings reveal a novel regulatory loop between miR-34a and KLF4 during keratinocytes replicative senescence. PMID: 29580988
- Knockdown of KLF4 promotes migration and invasion of non-small cell lung cancer (NSCLC) cells, whereas rescue of KLF4 expression reduces cell motility in miR25-overexpressing NSCLC cells. PMID: 29568911
- The subcellular localization of Klf4 might be related to the resistance of A549 cells to cisplatin. PMID: 29665649
- Authors observed that small activating RNAs (saRNAs) induced overexpression of KLF4 could promote cell migration/invasion in NCM460 and HCT116 cell lines. PMID: 29274293
- This is, to our knowledge, the first report of decreased expression of TFF3, SPDEF, KLF4, and goblet cell population in the colon of patients with Hirschsprung's disease (HSCR). Altered goblet cell function may result in intestinal barrier dysfunction contributing to the development of Hirschsprung-associated enterocolitis (HAEC). PMID: 29383490
- KLF4 overcomes tamoxifen resistance by suppressing the MAPK signaling pathway and predicts good prognosis in breast cancer. PMID: 28988130
- KLF4 activated the transcription activity of the iNOS promoter in MH7A cells stimulated by TNF-alpha. This study indicates that KLF4 is important for regulating the expression of iNOS by TNF-alpha in human synoviocytes. PMID: 28744810
- Anaplastic thyroid cancer cells show high expression of KLF4, and KLF4 expression is necessary for maintaining the undifferentiated phenotype and drug resistance. PMID: 28920531
- KLF4 could transcriptionally inhibit Cav-1 expression by binding directly to the promoter domain of Cav-1. PMID: 29587259
- Impairment of IGF2 gene expression in prostate cancer is triggered by epigenetic dysregulation of IGF2-DMR0 and its interaction with KLF4 PMID: 29017567
- Data show that Kruppel-like factor 4 (KLF4) was overexpressed in met proto-oncogene protein (c-Met)-overexpressing non-small-cell lung cancer (NSCLC) cells and tissues. PMID: 29624806
- KLF4 may act as an administered indicator to assess whether adjuvant postoperative pharmaceutical therapy is needed for patients with colorectal cancer. Low KLF4 expression was significantly correlated with reductions in overall survival and recurrence rate. PMID: 28752861
- Lower expression of KLF4 and NDRG2 in colorectal cancer patients was correlated with poor overall survival. Thus, KLF4 inhibited the proliferation of colorectal cancer cells dependent on NDRG2 signaling, which provides a novel strategy for therapy and early diagnosis of colorectal cancer. PMID: 28656310
- KLF4 is downregulated in anaplastic meningioma compared with low-grade meningioma subtypes. By manipulating KLF4 expression in anaplastic meningioma stem-like cells, this study demonstrated that KLF4 acts as a tumor suppressor during malignant progression in meningioma, affecting apoptosis, proliferation, invasion, and cell cycle. PMID: 28651379
- KLF4 was a direct target of miR-212, and miR-212 repressed KLF4 expression in a post-transcriptional manner. Moreover, miR-212-mediated protection effects were abated following KLF4 expression restoration in MPP-induced SH-SY5Y cells, represented as lowered cell viability and enhanced apoptotic rate. PMID: 29611404
- Results revealed that novel crosstalk between KLF4 and ZEB1 regulated gemcitabine resistance in pancreatic ductal adenocarcinoma (PDAC). PMID: 28849150
- This is the first demonstration that dysregulated KLF4 expression associates with poor differentiation of pancreatic cancer. Epigenetic activation of the miR-152/DNMT1/KLF4 signaling pathway by dietary DIM causes differentiation and significant growth inhibition of pancreatic cancer cells, highlighting its translational implications for pancreatic and other cancers. PMID: 28659310
- The present study found that the KLF4 and KLF5 3'-UTR contains one conserved target site of miR-506 and miR-124, and the overexpression of miR-506 and miR-124 inhibited the H2O2-induced upregulation of KLF4 and KLF5 in human coronary artery smooth muscle cells (HCMs). PMID: 28849090
- KLF4 enhances the sensitivity of cisplatin to esophageal squamous cell carcinoma (ESCC) cells through apoptosis induction and cell cycle arrest. PMID: 28694421
- KLF4 was identified as an important regulatory factor in the host response to fungi and as a controlling element in the IL-6 immune response with a unique expression pattern comparing fungal and lipopolysaccharide stimulation. PMID: 27346433
- KLF4 level in vascular endothelium decreased with age PMID: 29030550
- Results showed the expression levels of KLF4 were increased in bladder cancer (BC) patients which strongly correlated with the expression levels of PTBP1 but inversely correlated with the expression level of miR-145. These findings suggest that KLF4 is likely to positively contribute to carcinogenesis in certain types of BC. PMID: 28380435
- Transcriptional inhibition of MSI2 expression by KLF4 occurred in multiple PDAC cell lines as well as mouse models of PDAC. Lost expression of KLF4, a transcriptional repressor of MSI2 results in overexpression of MSI2 in PDACs, which may be a biomarker for accurate prognosis. A dysregulated KLF4/MSI2 signaling pathway promotes PDAC progression and metastasis. PMID: 27449499
- The results of the present study demonstrated that by regulating KLF4, miR145 may be involved in regulating smooth muscle differentiation of adipose-derived stem cells (ASCs) induced by TGFbeta1 and BMP4. PMID: 28440409
- Our findings reveal KLF4 as a key regulator of miR-182 cluster expression in human embryonic stem cells (hESCs) and a main contributor to its aberrant expression in melanoma and potentially in other tumors PMID: 28412746
- Surprisingly, 116 genes are directly activated via mCpG-dependent KLF4 binding activity. In-depth mechanistic studies reveal that recruitment of KLF4 to the methylated cis-regulatory elements of these genes results in chromatin remodeling and transcription activation. PMID: 28553926
- KLF4 transcriptionally repressed FOXO1 expression in glioma cells, contributing to glioma cell invasion and growth. PMID: 27835585
- Data suggest that KLF4 could promote cell senescence through a complex network: miR-203, survivin, and p21, which were all regulated by overexpression of KLF4 and contributed to cell senescence. PMID: 27531889
- Our results establish KLF4alpha as a KLF4 isoform that opposes the function of KLF4(FL) and as an important factor in the complex and unresolved role of KLF4(FL) in breast carcinogenesis. PMID: 27323810
- Kruppel-like factor 4 (KLF4) inactivation in chronic lymphocytic leukemia correlates with promoter DNA-methylation and can be reversed by inhibition of NOTCH signaling. PMID: 27081174
- Data suggest that the Kruppel-like factor 4 (KLF4) /telomerase reverse transcriptase (hTERT)/MAPK pathway is a potential new therapeutic target for lung cancer. PMID: 27153563
- The expression of KLF4 was significantly increased in human osteosarcoma tissues compared with the normal tissues. Elevated KLF4 promoted human osteosarcoma cell proliferation and metastasis. Mechanistic studies revealed KLF4 specifically bound the promoter of CRYAB and upregulated CRYAB expression in human osteosarcoma cells. PMID: 27105535
- In urothelial bladder carcinoma, strong KLF4 expression was associated with a higher risk of metastasis and death. KLF4 positively correlates with TWIST1 and vimentin, and inversely with E-cadherin. In vitro, it is accompanied by decreased E-cadherin and beta-catenin, increased vimentin and fibronectin, and enhanced migration/invasion. KLF4 knockdown suppressed TWIST1 expression and epithelial-mesenchymal transition (EMT), migration, and invasion. PMID: 27519276
- TGF-beta1 down-regulated KLF4 by activating miR-135a-5p, promoting proliferation and metastasis in hepatocellular carcinoma PMID: 27302923
- Our data suggest that KLF4 inhibits epithelial-mesenchymal transition (EMT)-enhanced hepatocellular carcinoma growth and invasion PMID: 27102441
- CSF and plasma expression of KLF4 mRNA was significantly decreased around 1-3 days after subarachnoid hemorrhage and remained lower than in controls. PMID: 28893694
- PARP1 recruits KLF4 to activate telomerase expression and stem cell pluripotency, indicating a positive regulatory role of the PARP1-KLF4 complex in telomerase expression in cancer and stem cells. PMID: 28985359
- KLF2 and KLF4 serve as important regulators that promote hemoglobin alpha expression in the endothelium. PMID: 28825355
- Loss of KLF4 expression is associated with T-cell acute lymphoblastic leukemia. PMID: 27872496
- IRF4 protects arteries against neointima formation by promoting the expression of KLF4 by directly binding to its promoter. PMID: 28851732
- High KLF4 expression is associated with breast cancer tumorigenesis. PMID: 28068319
- High KLF4 expression is associated with melanoma. PMID: 28068326
- KLF4 transactivates Ch25h and LXR, thereby promoting the synergistic effects between endothelial cells (ECs) and macrophages to protect against atherosclerosis susceptibility. PMID: 28794002
- Findings evidence a positive correlation between SIRT1 and BCL6 expression increase in follicular lymphomas (FL). SIRT1 methylation decreases in FL and diffuse large-B cell lymphomas (DLBCL), and this parallels the increase of KLF4, DAPK1, and SPG20 methylation. PMID: 28324774
Q&A
What is KLF4 and why is it significant in biological research?
KLF4 (Krüppel-like factor 4) functions as a transcription factor involved in cell proliferation, differentiation, and pluripotency maintenance. Its significance stems from its role as one of the crucial reprogramming factors in induced pluripotent stem cell generation. Research indicates that KLF4 may function either as a tumor suppressor or an oncogene depending on the tumor type, making it an important target in cancer research . In embryonic stem cells, KLF4 is a key regulator of pluripotency networks, while in differentiated tissues such as colon epithelium, it plays important roles in maintaining cellular identity .
What types of KLF4 antibodies are available for research applications?
Multiple types of KLF4 antibodies are commercially available, including:
These antibodies are derived from different immunogens and optimized for various experimental applications, allowing researchers to select the most appropriate tool for their specific research questions .
What is the expected molecular weight of KLF4 in Western blot analysis?
KLF4 typically appears at approximately:
The slight variation in observed molecular weight may result from post-translational modifications or differences in gel systems and sample preparation protocols across laboratories .
Which cell lines demonstrate reliable endogenous KLF4 expression?
Based on the available data, the following cell lines show consistent KLF4 expression and can serve as positive controls:
The HT-29 cell line in particular has been validated as a positive control across multiple antibody sources, while HDLM-2 (Human Hodgkin's Lymphoma Cell Line) serves as a negative control .
What are the recommended antibody concentrations for different applications?
Optimal concentrations vary by application and specific antibody:
Researchers should note that these values serve as starting points, and optimization for specific experimental conditions is recommended .
How can KLF4 antibodies be utilized in stem cell research?
KLF4 antibodies have become essential tools in stem cell biology research:
-
Pluripotency assessment: Detection of KLF4 expression in embryonic stem cells and iPSCs as a marker of stemness and pluripotency status
-
Reprogramming studies: Monitoring KLF4 expression during somatic cell reprogramming to assess conversion efficiency
-
Differentiation research: Tracking KLF4 expression changes during directed differentiation of stem cells
-
ChIP experiments: Identifying KLF4 genomic binding sites in pluripotent versus differentiated states
The search results demonstrate successful detection of KLF4 in BG01V human embryonic stem cells using immunofluorescence, showing its nuclear localization pattern characteristic of active transcription factors .
What insights can KLF4 antibodies provide about cancer biology?
KLF4 antibodies contribute significantly to cancer research through:
-
Expression profiling: Comparing KLF4 levels across various cancer cell lines (e.g., colorectal cancer lines HT-29, SW480, HCT-116)
-
Mechanistic studies: Investigating KLF4's dual role as either tumor suppressor or oncogene depending on cancer type
-
Pathway analyses: Examining interactions between KLF4 and other factors in cancer-related signaling pathways
-
Therapeutic response: Monitoring KLF4 expression changes following experimental treatments
Research has shown that KLF4 can be detected in the nuclei of epithelial cells in human colon tissue, providing insights into its normal expression pattern for comparison with cancer tissues .
How can ChIP assays with KLF4 antibodies identify gene regulatory networks?
Chromatin immunoprecipitation (ChIP) with KLF4 antibodies enables researchers to:
-
Map genomic binding sites: Identify promoter regions directly bound by KLF4 (e.g., B2R promoter)
-
Study transcriptional regulation: Analyze how KLF4 binding influences expression of target genes such as p21 and p27
-
Investigate epigenetic mechanisms: Examine interactions between KLF4 and chromatin modifiers like HDAC1
-
Compare binding profiles: Contrast KLF4 occupancy across different cell types or experimental conditions
The search results demonstrate successful ChIP experiments using anti-KLF4 antibodies to immunoprecipitate KLF4-DNA complexes, followed by PCR amplification of specific promoter regions .
What controls should be included when validating KLF4 antibody specificity?
Proper validation requires multiple controls:
-
Positive control cell lines: Include HT-29, SW480, or HCT-116 cells known to express KLF4
-
Negative control cells: Use HDLM-2 cells which show minimal KLF4 expression
-
Knockdown validation: Compare antibody signal between control and KLF4 knockdown samples (shKLF4)
-
Molecular weight verification: Confirm detection at the expected molecular weight (60-65 kDa)
-
Competing peptide: Where available, use the immunizing peptide to demonstrate binding specificity
The search results demonstrate knockdown validation where KLF4 antibody signal is significantly reduced in shKLF4 samples compared to control shRNA samples, confirming antibody specificity .
What are common challenges in KLF4 detection and how can they be addressed?
Researchers may encounter several technical challenges:
For Western blot applications, the search results indicate that using PVDF membrane under reducing conditions with appropriate buffer systems (e.g., Immunoblot Buffer Group 5) improves specificity .
How can KLF4 antibodies help elucidate interactions with cell cycle regulators?
KLF4 antibodies enable detailed investigation of cell cycle regulation mechanisms:
-
Expression correlation: Western blot analysis of KLF4 alongside cell cycle regulators like p21 and p27
-
Transcriptional control: ChIP assays to determine direct binding of KLF4 to cell cycle gene promoters
-
Functional studies: Examine effects of KLF4 knockdown or overexpression on cell cycle progression
-
Pathway interactions: Study how KLF4 interacts with other regulators like HDAC1 to control cell proliferation
Experimental data shows that KLF4 can significantly rescue the effects of HDAC1 on leukemia cell proliferation, suggesting a functional relationship between these factors in cell cycle control .
What role does KLF4 play in epithelial-to-mesenchymal transition (EMT)?
KLF4 antibodies have revealed important insights into EMT regulation:
-
Marker expression analysis: qRT-PCR and Western blot to correlate KLF4 levels with EMT markers
-
Functional studies: Compare mesenchymal marker expression (Fsp1, Id1) and endothelial markers (VE-cadherin, Claudin5) in KLF4 knockdown versus control cells
-
Signaling pathway interactions: Investigate how KLF4 influences stem cell-like characteristics (Sca1 expression) during EMT
The search results demonstrate that KLF4 knockdown in CCM1 KO endothelial cells significantly affects the expression of mesenchymal markers, suggesting KLF4 regulates the EndMT switch .
What techniques are recommended for quantifying KLF4 expression changes?
Multiple quantification approaches have been validated:
-
qRT-PCR: For mRNA expression analysis with fold-change calculation relative to control samples
-
Western blot densitometry: Quantifying protein levels with normalization to loading controls like GAPDH
-
Flow cytometry: For analyzing cell cycle effects in relation to KLF4 manipulation
-
Proliferation assays: Correlating cell proliferation (e.g., using CCK8 assay) with KLF4 expression levels
Statistical analysis should include appropriate tests (e.g., two-tailed unpaired t-test) with significance levels clearly indicated as demonstrated in the knockdown validation studies .
How should researchers interpret subcellular localization patterns of KLF4?
Interpretation of KLF4 localization should consider:
-
Nuclear localization: As a transcription factor, KLF4 primarily localizes to the nucleus in most cell types, particularly in epithelial cells of human colon tissue
-
Cytoplasmic staining: Some cytoplasmic KLF4 staining may be observed depending on cell type and state
-
Localization changes: Alterations in KLF4 localization may indicate changes in activity or regulatory status
-
Co-localization analysis: Association with other nuclear factors may provide functional insights
For accurate interpretation, counterstaining with nuclear markers (e.g., DAPI) is recommended as shown in the immunofluorescence protocols from the search results .
