FERMT1 Antibody, HRP conjugated
Description
Key Research Applications
The HRP-conjugated FERMT1 antibody facilitates precise detection in multiple experimental workflows:
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ELISA: Optimized for quantitative analysis of FERMT1 expression in biological samples .
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Western Blot: Validated for detecting FERMT1 at ~70–77 kDa in human and mouse tissues (e.g., kidney, HEK-293 cells) .
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Immunohistochemistry (IHC): Used to localize FERMT1 in formalin-fixed paraffin-embedded (FFPE) tissues, including cancer samples .
Western Blot Performance
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Positive Controls: Detected in human colon cancer cells (COLO 320), HEK-293 cells, and mouse kidney tissues .
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Band Specificity: A single band at ~77 kDa confirms minimal cross-reactivity .
IHC Staining
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Human Tissues: Strong staining observed in colon cancer, pancreatic cancer, and normal pancreas tissues .
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Protocol Notes: Antigen retrieval recommended using TE buffer (pH 9.0) or citrate buffer (pH 6.0) .
Supplier Landscape
A comparative analysis of suppliers offering FERMT1 HRP-conjugated antibodies:
Research Findings
FERMT1’s role in cancer progression (e.g., glioma, hepatocellular carcinoma) has been extensively studied using knockdown and functional assays . While the HRP-conjugated antibody itself is not directly cited in mechanistic studies, its application in validating FERMT1 expression has supported critical findings:
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Glioma: FERMT1 knockdown reduced proliferation, migration, and stemness via glycolytic pathway modulation .
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Hepatocellular Carcinoma (HCC): CARM1-mediated histone methylation upregulates FERMT1, promoting metastasis .
Protocol Recommendations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Kindlin-1 is primarily expressed in the cytoplasm of both normal esophageal squamous epithelium and esophageal cancer (EC) cells. Its expression is positively correlated with tumor cell differentiation and is elevated in stage I tumors. Notably, Kindlin-1 expression is higher in non-smoker patients compared to smoker patients and in individuals with a family history of EC. PMID: 28667517
- Kindlin supports platelet GPIIB IIIA activation by interacting with paxillin. PMID: 28954813
- Research indicates that Kindlin-1 promotes colorectal cancer (CRC) progression by recruiting SARA and Smad3 to TbetaRI, thereby activating the TGF-beta/Smad3 signaling pathway. This finding suggests that Kindlin-1 is a novel regulator of TGF-beta/Smad3 signaling and potentially a target for CRC therapeutics. PMID: 27776350
- Sequence analysis of KIND1 exons in a patient revealed a commonly reported homozygous nonsense mutation in exon 6 (c.811C>T;p.R271X). Additionally, two other patients exhibited novel homozygous single nucleotide deletions. PMID: 27862150
- Consistent follow-up care can effectively manage the periodontal disease activity in patients with Kindler syndrome (KS). PMID: 29168364
- Research has revealed a novel role for Kin1 in microtubule acetylation and stability. PMID: 26993041
- Keratinocytes derived from KS patients exhibit impaired electrotaxis. This defect can be restored by overexpressing wild-type Kindlin-1, but not by a W612A mutation that prevents Kindlin-integrin binding, highlighting the importance of Kindlin-integrin interaction in electrotaxis. PMID: 27427485
- FERMT1 activates beta-catenin transcriptional activity, thereby promoting epithelial-mesenchymal transition (EMT) in colorectal cancer metastasis. PMID: 27641329
- KIND1 plays a significant role in keratinocyte proliferation and suppressing UV-induced inflammation and DNA damage. PMID: 27725201
- Studies have shown a direct relationship between Kindlin-1 abundance and UV-B induced apoptosis in keratinocytes. Notably, Kindlin-2 overexpression does not have a compensatory effect. PMID: 27798104
- Evidence suggests that Kindlin-1 is crucial in EGF-induced re-epithelialization during skin wound healing. This finding provides additional rationale for the clinical application of EGF in treating acute wounds. PMID: 28290610
- KS is caused by mutations in the FERMT1 gene. Since 2003, over 60 FERMT1 mutations have been identified. While the FERMT1 mutation database is expanding, a clear genotype-phenotype correlation in KS remains elusive. PMID: 25865288
- A nonsense mutation in Exon 5 of the KIND1 gene identified in an Iranian family may lead to incomplete and non-functional protein products, suggesting pathogenicity and having significant implications for the diagnosis of patients with Kindler syndrome. PMID: 27293055
- Research reveals that some KS patients may harbor FERMT1 transcriptional regulatory mutations that are not routinely detected. PMID: 25156791
- Kindlin-1 is highly expressed in epithelial tissues derived from ectoderm and endoderm, whereas Kindlin-2 is predominantly expressed in mesoderm-derived tissues. Similarly, Kindlin-1 exhibits high expression in endoderm/ectoderm-derived tissues during embryonic development. PMID: 25591451
- FERMT1 mutation is a causative factor for Kindler syndrome. PMID: 26083552
- Data suggest that Kindlin-1 plays a crucial role in hepatocellular carcinoma and may serve as a promising prognostic marker and potential target for hepatocellular carcinoma therapy. PMID: 25592379
- A spectrum of FERMT1 mutations has been identified in 13 Iranian families diagnosed with Kindler syndrome. PMID: 25599393
- Researchers have identified a novel mutation in FERMT1. These findings align with the observation that the majority of KS-causing mutations in FERMT1 lead to premature termination of translation, resulting in loss of Kindlin-1 function in Kindler syndrome. PMID: 24635080
- The C-terminal LIM domains of migfilin determine its focal adhesion localization, and these domains mediate an interaction with Kindlin in vitro and in cells, demonstrating the importance of Kindlin for normal migfilin dynamics. PMID: 24165133
- Research has uncovered a role for Kindlin-1 in regulating integrin trafficking and adhesion turnover. PMID: 23776470
- Short interfering RNA-mediated depletion of Kindlin-1 leads to the formation of abnormal mitotic spindles. This effect depends on the ability of Kindlin-1 to bind integrins and Polo-like kinase 1-mediated Kindlin-1 phosphorylation. PMID: 23804033
- Individuals with Kindler syndrome (KS) exhibit loss-of-function mutations in the FERMT1 gene. PMID: 23278235
- While both Integrin-linked kinase (Ilk) and Kindlin-1 cooperate with Integrin alpha3beta1 to resist trauma-induced epidermal defects, surprisingly, Kindlin-1 and Ilk function in parallel, rather than synergistically. PMID: 23549420
- Kindlin-1 expression is implicated in the progression of pancreatic cancer through enhanced cell migration and invasion. PMID: 23440354
- Kindlin-1 and Kindlin-2 have opposing roles in lung cancers. PMID: 23209705
- Research suggests that FERMT1 is specifically expressed in colon carcinoma cells and plays roles in matrix invasion and cell growth. PMID: 23267142
- Direct sequencing of the FERMT1 gene revealed a homozygous insertion of cytosine at position 676 (c.676insC) in exon 5 in seven patients. PMID: 22220914
- There is an association of FERMT1 missense and in-frame deletion mutations with milder disease phenotypes and a later onset of complications in Kindler syndrome (FERMT1). PMID: 21936020
- Kindlin-1 expression in breast tumors is associated with lung metastasis and lung metastasis-free survival through the regulation of TGF-beta signaling. Silencing Kindlin-1 prevented tumor growth and lung metastasis in mice. PMID: 21832234
- FERMT1 is a novel prognostic factor for colon carcinoma. PMID: 21220475
- This study describes five novel and three recurrent loss-of-function FERMT1 mutations in eight individuals with Kindler syndrome, providing an overview of genotype-phenotype correlation in this disorder. PMID: 21336475
- FERMT1 mutations induce phenotype-modifying cytokines. PMID: 21309038
- The phenotype of Kindlin-1-deficient cells can be modulated by regulating Kindlin-2 gene expression and vice versa. PMID: 21356350
- This research describes a recurrent splice-site deletion mutation in KIND1 in Kindler syndrome. PMID: 21146372
- A novel mutation in the FERMT1 gene in a Spanish family with Kindler's syndrome is reported. PMID: 20028441
- This review discusses the cellular functions and potential clinical relevance of Kindlin-1. PMID: 19854292
- Null mutations in FERMT1 result in skin blistering from birth, early childhood progressive poikiloderma, mucosal fragility, and an increased risk of cancer. PMID: 19945623
- Kindlin plays a role in mediating cell processes that depend on integrins. PMID: 14634021
- Loss-of-function KIND1 mutations demonstrate the importance of Kindlin-1 in maintaining epithelial integrity. PMID: 14962093
- Kindlin-1 is considered a component in the linkage of the actin cytoskeleton to the extracellular matrix, suggesting both structural and cell-signaling functions. This is a review article. PMID: 15927810
- Mutated at intron 13 in Kindler syndrome. PMID: 16051467
- The abundance of repetitive elements in intronic regions of KIND1, along with the identification of a large deletion, suggests that genomic rearrangements could be responsible for a significant proportion of Kindler syndrome cases. PMID: 16675959
- Kindlin-1 plays roles in the regulation of polarity, proliferation, and motility of epidermal keratinocytes. PMID: 17012746
- Kindlin-1 connects the actin cytoskeleton to the extracellular matrix and is believed to have cell-signaling functions due to its various functional domains. PMID: 17178989
- This research analyzes KIND1 gene mutations in Kindler syndrome, presenting case reports. PMID: 17460733
- The KIND1 mutation c.67insC is the most common recurrent pathogenic gene mutation observed in patients with KS. PMID: 17916195
- Two patients with Kindler Syndrome have mutations in KIND-1. In patient 1, a duplication of cytosine at position 676 in exon 5 of Kindlin-1 mRNA was identified. In patient 2, a novel mutation in exon 3 of the KIND1 gene (c.170C>A) was found. PMID: 17989907
- A splice site mutation in the first position of intron 13 of the FERMT1 gene leads to the skipping of exon 13. PMID: 18652585
- A novel large FERMT1 (KIND1) gene deletion in Kindler syndrome has been reported. PMID: 18835760
Q&A
What is FERMT1 and why is it a significant research target?
FERMT1 is a FERM domain-containing adaptor protein predominantly found at cell-extracellular matrix adhesions where it binds to β-integrin subunits and is required for integrin activation. It plays crucial roles in cell adhesion, keratinocyte proliferation, normal polarization of basal keratinocytes in skin, and cell migration to wound sites . FERMT1 has become a significant research target as it's highly expressed in many tumors, including non-small cell lung cancer (NSCLC), nasopharyngeal carcinoma (NPC), and colon carcinoma, where it acts as an oncogene promoting cell migration and invasion .
How should FERMT1 antibody, HRP conjugated be stored for optimal stability?
For optimal stability, HRP-conjugated FERMT1 antibody should be shipped at 4°C and upon receipt stored at either -20°C for short-term storage or -80°C for long-term storage . Repeated freeze-thaw cycles should be avoided to maintain antibody integrity and functionality. Some preparations are supplied in buffers containing 50% glycerol, 0.01M PBS (pH 7.4), and 0.03% Proclin 300 as preservatives , which helps maintain stability during storage.
Which tissue and cell types show positive reactivity with FERMT1 antibody?
FERMT1 antibody has demonstrated positive reactivity in both human and mouse samples . Specifically, Western blot applications have successfully detected FERMT1 in mouse kidney tissue, COLO 320 cells, and HEK-293 cells. Immunohistochemistry applications have shown positive results in human colon cancer tissue, human pancreas tissue, and human pancreas cancer tissue. Immunofluorescence applications have detected FERMT1 in HEK-293 cells . These validated tissue and cell types provide researchers with reliable positive controls for experimental design.
What are the optimal antigen retrieval methods when using FERMT1 antibody in IHC applications?
For immunohistochemistry applications with FERMT1 antibody, the recommended antigen retrieval method is using TE buffer at pH 9.0. Alternatively, citrate buffer at pH 6.0 may also be used for antigen retrieval . When performing IHC with FERMT1 antibody on formalin-fixed paraffin-embedded tissues, these antigen retrieval steps are critical for unmasking epitopes that may be crosslinked or obscured during fixation. Proper antigen retrieval significantly improves staining intensity and specificity, particularly in tissues with complex extracellular matrix components where FERMT1 is typically localized.
How can researchers validate knockdown efficiency in FERMT1 functional studies?
To validate FERMT1 knockdown efficiency, researchers should employ a combination of techniques. Western blotting and real-time PCR are commonly used to detect the efficiency of transfection following shRNA introduction . For shRNA-mediated knockdown, multiple target sequences have been validated in previous studies, including:
Proper validation using both protein and mRNA detection methods ensures reliable interpretation of subsequent functional assays investigating FERMT1's role in cellular processes.
What controls should be included when using FERMT1 antibody, HRP conjugated in ELISA?
When using HRP-conjugated FERMT1 antibody in ELISA, researchers should include several critical controls:
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Positive control: Lysates from cells known to express FERMT1 (e.g., HEK-293 cells, COLO 320 cells)
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Negative control: Samples from FERMT1 knockout cells or tissues
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Antibody specificity control: Pre-absorption with the immunizing peptide
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Background control: Wells without primary antibody but with all other reagents
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Standard curve: Using recombinant FERMT1 protein for quantitative analysis
These controls help validate assay specificity, sensitivity, and reproducibility, ensuring reliable FERMT1 detection and quantification in experimental samples.
How can FERMT1 antibody be utilized to investigate epithelial-mesenchymal transition (EMT) in cancer research?
FERMT1 has been shown to regulate epithelial-mesenchymal transition (EMT) in multiple cancer types, making FERMT1 antibody a valuable tool for EMT research . To investigate this:
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Use FERMT1 antibody in conjunction with EMT markers (E-cadherin, N-cadherin, Vimentin) in Western blot or immunofluorescence to correlate FERMT1 expression with EMT status
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Perform FERMT1 knockdown or overexpression followed by detection of EMT markers to establish causality
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Implement wound healing and Transwell assays after modulating FERMT1 expression to assess functional consequences on migration and invasion
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Co-stain for FERMT1 and PKP3 in immunofluorescence studies, as FERMT1 has been shown to upregulate PKP3 expression to promote invasion and migration
This comprehensive approach helps delineate the specific role of FERMT1 in driving EMT programs across different cancer contexts.
What is the relationship between FERMT1 and the p38 MAPK signaling pathway, and how can researchers investigate this connection?
FERMT1 has been shown to activate the p38 MAPK signaling pathway through upregulation of PKP3 expression . To investigate this signaling connection:
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Use Western blotting to detect phosphorylated p38 MAPK levels following FERMT1 overexpression or knockdown
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Perform co-immunoprecipitation (Co-IP) assays to identify direct or indirect interactions between FERMT1 and components of the MAPK pathway
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Employ specific p38 MAPK inhibitors in FERMT1-overexpressing cells to determine if migration and invasion phenotypes are dependent on p38 MAPK activity
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Conduct dual knockdown experiments targeting both FERMT1 and PKP3 to validate the proposed pathway mechanism
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Use phospho-specific antibodies against p38 MAPK pathway components in combination with FERMT1 antibody in immunofluorescence to visualize pathway activation in specific cellular compartments
This systematic approach will help establish the mechanistic details of how FERMT1 regulates MAPK signaling in cancer progression.
How can gene expression profiling be combined with FERMT1 antibody studies to identify novel downstream targets?
Integrating gene expression profiling with FERMT1 antibody studies provides a powerful approach to identify novel downstream targets:
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Perform RNA-seq or microarray analysis on FERMT1-overexpressing or FERMT1-knockdown cells
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Use Gene Set Enrichment Analysis (GSEA) to identify significantly enriched pathways, as previously demonstrated in FERMT1 studies
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Validate protein-level changes of candidate genes using Western blotting with FERMT1 antibody as a control
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Analyze public databases (such as GEPIA and TCGA) to identify genes positively correlated with FERMT1 expression in cancer tissues, as previously done to identify the FERMT1-PKP3 correlation
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Perform chromatin immunoprecipitation (ChIP) assays to determine if FERMT1 directly or indirectly regulates transcription of candidate genes
This integrated approach can reveal previously unidentified FERMT1-regulated genes and pathways relevant to cancer progression.
What are common issues when using FERMT1 antibody in Western blotting and how can they be resolved?
Several common issues may arise when using FERMT1 antibody in Western blotting:
Using validated positive control samples such as HEK-293 or COLO 320 cell lysates can help establish appropriate experimental conditions and troubleshoot detection issues .
How can researchers optimize immunohistochemistry protocols for FERMT1 detection in different tissue types?
Optimizing immunohistochemistry protocols for FERMT1 detection requires tissue-specific considerations:
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Fixation optimization: Use 10% neutral buffered formalin for 24-48 hours; excessive fixation can mask epitopes
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Antigen retrieval methods: Compare TE buffer (pH 9.0) versus citrate buffer (pH 6.0) as different tissues may respond better to specific retrieval methods
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Antibody dilution titration: Test multiple dilutions (1:50, 1:100, 1:200, 1:500) to determine optimal concentration for each tissue type
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Incubation conditions: Optimize primary antibody incubation time (4°C overnight versus 1-2 hours at room temperature)
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Detection systems: Compare avidin-biotin complex (ABC) versus polymer-based detection systems for best signal-to-noise ratio
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Counterstaining: Adjust hematoxylin intensity to maintain visibility of FERMT1 staining while providing adequate nuclear detail
Researchers should validate their protocol using known positive control tissues such as human colon cancer tissue or human pancreas tissue, where FERMT1 expression has been confirmed .
What considerations should be taken when designing multiplexed assays involving FERMT1 antibody?
When designing multiplexed assays with FERMT1 antibody, researchers should consider:
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Antibody species compatibility: Choose primary antibodies raised in different host species to avoid cross-reactivity
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Fluorophore selection: If using fluorescent detection, select fluorophores with minimal spectral overlap
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Epitope accessibility: Consider whether multiple antibodies targeting closely related proteins might cause steric hindrance
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Sequential staining: For challenging combinations, implement sequential rather than simultaneous staining protocols
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Validation controls: Include single-stain controls to verify specificity in the multiplexed context
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Cross-blocking experiments: Test whether one antibody blocks binding of another in the multiplex panel
For co-localization studies with FERMT1, consider combining with antibodies against integrin subunits, PKP3, or components of the MAPK pathway based on known interactions .
How does FERMT1 expression vary across different cancer types and what are the implications for antibody detection methods?
Research has demonstrated variable FERMT1 expression across different cancer types:
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Non-small cell lung cancer (NSCLC): Significantly upregulated FERMT1 expression correlates with poor prognosis
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Nasopharyngeal carcinoma (NPC): Elevated FERMT1 expression inhibits EMT and induces cell cycle arrest
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Colon carcinoma: Cancer cell-specific expression of FERMT1 enhances invasive ability and cell growth
These expression patterns have important implications for antibody detection methods:
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Tissue-specific optimization of antibody dilutions is necessary (starting with 1:50-1:500 for IHC)
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Cancer-specific positive controls should be selected based on known expression patterns
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Quantitative methods like Western blotting with densitometry analysis may be required to detect subtle differences in expression levels between cancer types
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Antibody validation should include multiple cancer cell lines to confirm specificity across different tumor contexts
Understanding these cancer-specific expression patterns helps researchers properly design and interpret FERMT1 antibody-based experiments.
What experimental approaches can investigate the interaction between FERMT1 and integrin activation in cancer cell metastasis?
To investigate FERMT1's role in integrin activation during cancer metastasis, researchers can employ:
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Co-immunoprecipitation (Co-IP): To detect direct interactions between FERMT1 and β-integrin subunits
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Proximity ligation assay (PLA): To visualize and quantify FERMT1-integrin interactions in situ
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FRET/FLIM analysis: To measure dynamics of FERMT1-integrin binding in living cells
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Adhesion assays: Compare adhesion to extracellular matrix proteins (fibronectin, laminin) in FERMT1-modulated cells
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Live-cell imaging: Track integrin clustering and focal adhesion formation in relation to FERMT1 expression
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3D invasion assays: Evaluate the impact of FERMT1 knockdown on directional migration through extracellular matrix
These approaches provide complementary insights into how FERMT1 contributes to integrin activation and subsequent metastatic capabilities in cancer cells.
How can FERMT1 antibody be used to evaluate potential therapeutic targets in the FERMT1 signaling pathway?
FERMT1 antibody can be instrumental in evaluating therapeutic targets within its signaling pathway:
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Target validation: Use FERMT1 antibody in Western blotting to confirm knockdown efficacy of potential therapeutic approaches targeting FERMT1
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Pathway monitoring: Track changes in downstream effectors like PKP3 and phospho-p38 MAPK following treatment with pathway inhibitors
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Combination therapy assessment: Evaluate FERMT1 expression and activation status when combining p38 MAPK inhibitors with other targeted therapies
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Patient stratification biomarker development: Develop IHC protocols using FERMT1 antibody to identify patients likely to respond to therapies targeting the FERMT1-PKP3-MAPK axis
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Resistance mechanism studies: Monitor FERMT1 expression changes in cell lines developing resistance to targeted therapies
These applications position FERMT1 antibody as a valuable tool in translational research aimed at developing new therapeutic approaches for cancers with FERMT1 pathway activation.
