Fas Ligand Rabbit Polyclonal Antibody
cat.: ER1908-69
Product Type: Rabbit polyclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Tissue, FC
Clonality: Polyclonal
Form: Liquid
Storage condition: Store at -20℃ for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20℃. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4℃.
Storage buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: 31 KDa
Isotype: IgG
Immunogen: KLH conjugated synthetic peptide derived from human Fas Ligand 196-281/281
Subcellular location: Cell membrane; Single-pass type II membrane protein. Secreted. Cytoplasmic vesicle lumen. Lysosome lumen. Note=May be released into the extracellular fluid, probably by cleavage form the cell surface. Is internalized into multivesicular bodies of secretory lysosomes after phosphorylation by FGR and monoubiquitination.
Recommended Dilutions:
  WB
  IHC-P
  IF-tissue
  FC
1:500-2000
1:100-500
1:100-500
1μg/Test
Uniprot #: SwissProt: P48023 Human
Alternative names: ALPS1B Apoptosis (APO 1) antigen ligand 1 Apoptosis antigen ligand 1 Apoptosis antigen ligand APT1LG1 APTL CD178 CD178 antigen CD95 ligand CD95-L CD95L CD95L protein Fas antigen ligand Fas L Fas ligand (TNF superfamily member 6) Fas ligand FASL Fasl Fas ligand (TNF superfamily member 6) FASLG Generalized lymphoproliferative disease Gld soluble form TNFL6_HUMAN TNFSF6 Tumor necrosis factor (ligand) superfamily member 6 Tumor necrosis factor ligand superfamily member 6
Images
ER1908-69_1.jpg Fig1: Sample:
Jurkat(Human) Cell Lysate at 30 ug
Primary: Anti-Fas Ligand (bs-0216R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kD
Observed band size: 40 kD
ER1908-69_2.jpg Fig2: Sample: Bone (mouse) Lysate at 40 ug
Primary: Anti- Fas ligand (bs-0216R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kD
Observed band size: 46 kD
ER1908-69_3.jpg Fig3: Sample:
Lane 1: Bone (Mouse) Lysate at 40 ug
Lane 2: Bone (Rat) Lysate at 40 ug
Primary: Anti-Fas Ligand (bs-0216R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 40 kD
Observed band size: 40 kD
ER1908-69_4.jpg Fig4: Tissue/cell: rat brain 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-FasL Polyclonal Antibody, Unconjugated(bs-0216R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1908-69_5.jpg Fig5: Tissue/cell: human rectal carcinoma; 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-FasL Polyclonal Antibody, Unconjugated(bs-0216R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining
ER1908-69_6.jpg Fig6: Paraformaldehyde-fixed, paraffin embedded (mouse placenta); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Fas Ligand) Polyclonal Antibody, Unconjugated (bs-0216R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
ER1908-69_7.jpg Fig7: Tissue/cell: human rectal carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-FasL Polyclonal Antibody, Unconjugated(bs-0216R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
ER1908-69_8.jpg Fig8: Blank control: mouse thymouses(blue)
Isotype Control Antibody: Rabbit IgG(orange) ;
Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue),
Dilution: 1:100 in 1 X PBS containing 0.5% BSA ;
Primary Antibody Dilution: 1μl in 100 μL1X PBS containing 0.5% BSA(green).
ER1908-69_9.jpg Fig9: Blank control: Mouse Kidney (blue). Primary Antibody:Rabbit Anti-phospho-Fas Ligand antibody (bs-0216R,Green); 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-FITC(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.

Protocol
The cells were fixed with 2% paraformaldehyde for 10 min at 37℃. Primary antibody (bs-0216R, 1μg /1x10^6 cells) were incubated for 30 min at room temperature, 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/FITC antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 40 min on ice. Acquisition of 20,000 events was performed.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.