Fas Ligand Rabbit Polyclonal Antibody

• Catalog Number: orb221406
• Category: Antibodies
• Description: Fas Ligand Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Fas Ligand (196-281/281aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 38-42 kDa
• Target: FASLG
• UniProt ID: P48023
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Tumor necrosis factor ligand superfamily member 6;
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: Mouse Kidney (blue). Primary Antibody: Rabbit Anti-phospho-Fas Ligand antibody (orb221406, Green), dilution: 1 µg 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 1X PBS containing 0.5% BSA. Protocol, The cells were fixed with 2% paraformaldehyde for 10 min at 37°C. Primary antibody (orb221406, 1 µg/1x10^6 cells) were incubated for 30 min at room temperature, followed by 1X 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 20000 events was performed.

Blank control: mouse thymouses (blue), Isotype Control Antibody: Rabbit IgG (orange), Secondary Antibody: Goat anti-rabbit IgG-FITC (white blue), dilution: 1:100 in 1X PBS containing 0.5% BSA, Primary Antibody, dilution: 1 µl in 100 µl 1X PBS containing 0.5% BSA (green).

Paraformaldehyde-fixed, paraffin embedded (mouse placenta), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (Fas Ligand) Polyclonal Antibody, Unconjugated (orb221406) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Bone (mouse) Lysate at 40 ug, Primary: Anti-Fas ligand (orb221406) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 31 kD, Observed band size: 46 kD.

Sample: Jurkat (Human) Cell Lysate at 30 ug, Primary: Anti-Fas Ligand (orb221406) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 31 kD, Observed band size: 40 kD.

Sample: Lane 1: Bone (Mouse) Lysate at 40 ug, Lane 2: Bone (Rat) Lysate at 40 ug, Primary: Anti-Fas Ligand (orb221406) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 40 kD, Observed band size: 40 kD.

Tissue/Cell: human rectal carcinoma, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-FasL Polyclonal Antibody, Unconjugated (orb221406) 1:200, overnight at 4°C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated (orb868589) used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei.