Fas, also known as CD95 and TNFRSF6, is a 45kD type I transmembrane glycoprotein. The ECD contains two cysteine-rich domains, and the cytoplasmic tail contains one pro-apoptotic death domain. Alternate splicing generates five additional isoforms of feline Fas, all of which lack the transmembrane segment. Fas binding to Fas Ligand triggers apoptosis through both the MEK cascade and FADD/caspase-8 pathway. Within the ECD feline Fas shares 68%, 65%, 53%, and 58% aa identity with human, mouse, porcine, and rat Fas, respectively. Feline PBMC activated with PMA plus Ca 2+ ionomycin were stained with C2442-30D or isotype control followed by PE-conjugated anti-mouse antibody
Applications
Suitable for use in Flow cytometry, Western blot and Direct ELISA . Other applications have not been tested.
Recommended Dilutions
Flow cytometry: 25 ug/ml and add 10il of the diluted solution to 1-2.5 x 10e5 cells in a total reaction volume not exceeding 200ul. The binding of unlabeled monoclonal antibodies may be visualized by adding a secondary developing reagent such as goat anti-mouse IgG conjugated to a fluorochrome. Western Blot: 1-2ug/ml. the detection limit for rfeFas is approximately 25 ng/lane under non-reducing conditions. It is not recommended to use this antibody under reducing conditions. Chemiluminescent detection with WesternGloTM Chemiluminescent Detection Substrate will increase sensitivity by 5 to 50 fold. Direct ELISA: 0.5-1.0ug/ml The detection limit for rfeFas is approximately 3 ng/well. Optimal dilutions to be determined by the researcher.
Storage and Stability
Lyophilized and reconstituted products are stable for 12 months after receipt at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Immunogen
NS0-derived recombinant feline rfeFas extracellular domain.
Form
Supplied as a lyophilized powder from PBS, 5% trehalose. Reconstitute with 200ul sterile PBS.
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes feline Fas extracellular domain. Species sequence homology: human 68%, mouse 65%, porcine 53%, and rat 58%. Does not crossreact with rhFas, rmFas, or rrFas.
TDS
NCBI
UniProt
Tech Support
Customer Service
