RNF6 Antibody, FITC conjugated
Description
Structure and Conjugation
RNF6 Antibody, FITC conjugated, combines a primary antibody (polyclonal or monoclonal) specific to RNF6 with fluorescein isothiocyanate (FITC), a fluorescent dye. The conjugation occurs via primary amine groups (e.g., lysine residues) on the antibody, typically yielding 3–6 FITC molecules per antibody to avoid solubility issues or quenching .
Key Features of FITC Conjugation
Applications in Research
The RNF6 Antibody, FITC conjugated, is primarily used in fluorescence-based techniques to study RNF6 localization and function.
Common Techniques
RNF6 Protein and Its Role in Cancer
RNF6 is overexpressed in multiple cancers, including gastric, colorectal, breast, and hepatocellular carcinoma (HCC), where it promotes tumorigenesis, metastasis, and drug resistance .
RNF6-Driven Pathways
Antibody Specificity and Validation
RNF6 antibodies are validated for specificity and cross-reactivity across species.
Comparison of RNF6 Antibodies
Conjugation Protocols and Challenges
The LYNX Rapid Fluorescein Antibody Conjugation Kit (Bio-Rad) is widely used for FITC labeling .
Stepwise Protocol
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Antibody Preparation: Use 10–200 µg antibody in amine-free buffer (1 mg/mL, pH 6.5–8.5) .
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Conjugation Reaction: Mix antibody with lyophilized FITC reagent and modifier/quencher. Incubate at room temperature for 30–60 minutes .
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Purification: Remove unbound FITC using spin columns or dialysis .
Critical Notes
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Sodium Azide Removal: Essential to prevent FITC inactivation .
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Concentration Optimization: Higher FITC ratios (>6) may reduce antibody solubility .
Case 1: RNF6 in Gastric Cancer
Knockdown of RNF6 in gastric cancer cells (AGS, HGC-27) reduced STAT3 phosphorylation and increased SHP-1 levels, inhibiting cell growth and enhancing doxorubicin sensitivity . FITC-conjugated antibodies could localize RNF6 in these models.
Case 2: RNF6 in Colorectal Cancer
RNF6 overexpression in CRC cells upregulates SF3B2, a splicing factor, promoting tumorigenesis. FITC-labeled antibodies could track RNF6 nuclear localization during transcriptional activation .
Therapeutic Implications
Targeting RNF6-SF3B2 or RNF6-YAP axes with inhibitors (e.g., pladienolide B) shows promise in CRC and breast cancer models . FITC-conjugated antibodies aid in validating RNF6 as a therapeutic biomarker.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- High RNF6 expression has been linked to breast cancer. PMID: 28223545
- Research suggests that RNF6 overexpression in leukemia is driven by PBX1, indicating the potential for targeting the PBX1/RNF6 axis as a novel therapeutic strategy for leukemia. PMID: 26971355
- Studies have shown that Rnf6 regulates cellular LIMK1 concentrations, highlighting a novel function for the ubiquitin/proteasome system in regulating local growth cone actin dynamics. PMID: 16204183
- Research has identified and characterized a novel RNF6 transcriptional splice variant, Spg2, in human development. PMID: 18368307
- Functional analysis of the mouse homolog has been conducted. PMID: 11971979
