Western blot analysis of extracts from various cell lines using C9orf7/CACFD1 Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 263
C9orf7/CACFD1 Antibody recognizes endogenous levels of total C9orf7/CACFD1 protein. Based on the antigen sequence, this antibody is predicted to detect isoforms 1 and 2 of C9orf7/CACFD1 protein, but not isoforms 3 and 4.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu168 of human C9orf7/CACFD1 protein.
C9orf7/Calcium Channel Flower Domain Containing 1 (CACFD1), also known as Flower (FWE), is the human homolog of the Drosophila fwe gene. In Drosophila, expression of specific FWE membrane protein isoforms indicates the fitness of a cell during clonal selection and cell competition processes, which promotes the elimination of weaker cells from a growing population (1). In humans, there are four isoforms of C9orf7/CACFD1. Investigators have shown expression of human C9orf7/CACFD1 isoforms 1 and 3 indicates reduced cellular fitness compared to cells expressing isoforms 2 and 4, which provide a competitive advantage in survival. Moreover, human cancer cells show increased expression of isoforms 2 and 4 that confers a competitive growth advantage for cancer cells. Inhibition of C9orf7/CACFD1 expression reduces tumor growth and metastasis, and induces sensitivity to chemotherapy (2-5).
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