WFIKKN1 Antibody, FITC conjugated
Description
Functional Applications
Flow Cytometry:
Demonstrated in A549 cell line studies using FITC-conjugated secondary antibodies, achieving specific detection with signal-to-noise ratios >10:1 when using proper isotype controls . The 488 nm laser line excitation matches FITC's spectral properties for optimal detection in standard flow cytometers .
Western Blot:
Detects ~72 kDa FITC-BSA conjugates at concentrations as low as 0.1 ng/μL when using enhanced chemiluminescence substrates . The FITC moiety allows dual detection through both fluorescence and chemiluminescent methods.
Protein Interaction Studies:
WFIKKN1's functional domains show specific binding interactions:
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Follistatin domain binds mature GDF11/myostatin (Kd = 2.4 nM)
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NTR domain interacts with myostatin propeptide (Kd = 8.7 nM)
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Full-length WFIKKN1 exhibits 3x higher affinity than isolated domains
Validation Data
Specificity Controls:
Performance Metrics:
| Application | Sensitivity | Dynamic Range | CV% (Inter-assay) |
|---|---|---|---|
| Flow Cytometry | 100 cells/μL | 10^2-10^5 | <8% |
| Western Blot | 0.1 ng | 0.1-100 ng | <12% |
| ELISA | 50 pg/mL | 50-10^4 pg/mL | <15% |
Data compiled from manufacturer specifications and independent validation studies .
Experimental Considerations
Optimization Guidelines:
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Titrate between 1:50-1:500 dilution for flow cytometry
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Use 0.1% BSA in wash buffers to reduce non-specific binding
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Protect from light exposure >30 minutes during experiments
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Pair with anti-FITC secondary antibodies for signal amplification
Limitations:
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- This review summarizes our current understanding of WFIKKN proteins and proposes that they function as "companion" proteins for various growth factors. By providing localized and sustained presentation of TGFβ proteins to their respective receptors, they regulate the balance between the activation of Smad and non-Smad pathways by TGFβ. PMID: 27325460
- Research indicates that the KKN1 fragment generated by BMP1-cleavage of WFIKKN1 protein significantly contributes to the observed enhancer activity. PMID: 27782377
- Data suggest that WFIKKN1, but not WFIKKN2, can block myostatin binding to its receptor (activin receptors type II). Moreover, various forms of myostatin (promyostatin, myostatin pro-domain, and latent/processed myostatin) bind to WFIKKN1 but not WFIKKN2. PMID: 23829672
- Findings indicate that WFIKKN proteins act as antagonists of GDF8 and GDF11. However, in the case of TGFbeta1, BMP2, and BMP4, they function as growth factor binding proteins. PMID: 21054789
- This study compares the newly discovered WFIKKNPR gene structure and tissue distribution of gene expression with that of the WFIKKN gene in adult and fetal tissues and organs. PMID: 11928817
- Data suggest that the physiological target of WFIKKN protein may be trypsin. PMID: 12709070
- This study examines the three-dimensional structure of a Kunitz domain with a tryptophan residue at the P02 site of the protease-recognition loop. PMID: 16791741
- Both WFIKKN1 and WFIKKN2 exhibit high affinity for growth and differentiation factors 8 and 11. PMID: 18596030
