Product nameHuman FAS knockout HeLa cell lysate
See all Fas kits
Cell line information
Parental cell line: HeLa
Organism: Homo sapiens
Gene editing information
Editing tool: CRISPR/Cas9
Mutation: 1 bp insertion in exon2.
Knockout validation: Confirmed by Sanger sequencing and Western blot.
Reconstitution instructions: To use as a WB control, resuspend in 45 µL of Sample buffer (40% (w/v) Glycerol, 4% (w/v) Lithium Dodecyl Sulfate, 4% Ficoll 400, 0.025% Phenol Red, 0.025% Brilliant Blue G250, 2 mM EDTA) and 5 µL of DTT to resuspend @ 2mg/ml. Mix well, then boil the sample for 10 minutes before loading it onto the gel.
User storage instructions: Upon receiving, lysate can be diluted with 1 x SDS sample buffer & will be stable at -20°C for 12 months. Long term storage at -80°C.
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit Human FAS knockout HeLa cell lysate (Lyophilized) 1 x 100µg Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
FunctionReceptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS-mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both. The secreted isoforms 2 to 6 block apoptosis (in vitro).
Tissue specificityIsoform 1 and isoform 6 are expressed at equal levels in resting peripheral blood mononuclear cells. After activation there is an increase in isoform 1 and decrease in the levels of isoform 6.
Involvement in diseaseDefects in FAS are the cause of autoimmune lymphoproliferative syndrome type 1A (ALPS1A) [MIM:601859]; also known as Canale-Smith syndrome (CSS). ALPS is a childhood syndrome involving hemolytic anemia and thrombocytopenia with massive lymphadenopathy and splenomegaly.
Sequence similaritiesContains 1 death domain.
Contains 3 TNFR-Cys repeats.
DomainContains a death domain involved in the binding of FADD, and maybe to other cytosolic adapter proteins.
Cellular localizationSecreted and Cell membrane.
- Information by UniProt
- ALPS 1A
- APO 1
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: FAS knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - ab133619 observed at 37 kDa. Red - loading control ab8245 observed at 37 kDa.
ab133619 Anti-Fas antibody [EPR5700] was shown to specifically react with Fas in wild-type HeLa cells in western blot. Loss of signal was observed when knockout sample ab256911 was used. Wild-type and Fas knockout samples were subjected to SDS-PAGE. ab133619 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Homozygous: 1 bp insertion in exon2
ab256911 has not yet been referenced specifically in any publications.