Anti-CD95/FAS antibody (bs-0215R)

Anti-CD95/FAS antibody 

DATASHEET

Host:Rabbit

Target Protein:CD95/FAS

IR:Immunogen Range:35-110/327

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:246097

Swiss Prot:P25446

Source:KLH conjugated synthetic peptide derived from rat FAS:35-110/327 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background:FAS is a receptor for TNFSF6/FASL. The adaptor 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. Apoptosis or programmed-cell death is a physiological process essential for the normal development and maintenance of homeostasis in many organisms. This “cellular suicide” can be mediated by the Fas antigen (CD95, APO1), a cell-surface glycoprotein, 40-50kDa, that belongs to the nerve growth factor/tumor necrosis factor (TNF) receptor family. FAS-mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both (By similarity). It is type I membrane protein. Contains a death domain involved in the binding of FADD, and maybe to other cytosolic adaptor proteins Contains 1 death domain.

Size:100ul

Concentration:1mg/ml

Applications:ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1μg/Test)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Tissue/cell: rat pancreas tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Fas Polyclonal Antibody, Unconjugated(bs-0215R) 1:600, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell: rat colon tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Fas Polyclonal Antibody, Unconjugated(bs-0215R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control: Hela(blue).
Primary Antibody:Rabbit Anti-CD95FAS antibody(bs-0215R), Dilution: 1μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG(orange) ,used under the same conditions );
Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde (10 min). Antibody (bs-0215R, 1μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody of bs-0215R at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.