Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,Mouse,Rat (predicted:Cow)

IgG

Human,Mouse,Rat (predicted:Cow)

1. Sample:
Jurkat (Human) Cell Lysate at 30 μg
Primary: Anti-Fas Ligand (TMAB-00655) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 40 kDa
2. Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-FasL Polyclonal Antibody, Unconjugated (TMAB-00655) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Tissue/cell: human rectal carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-FasL Polyclonal Antibody, Unconjugated (TMAB-00655) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Tissue/cell: human rectal carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-FasL Polyclonal Antibody, Unconjugated (TMAB-00655) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated used at 1:200 dilution for 40 minutes at 37°C. DAPI (5 μg/ml,blue) was used to stain the cell nucleus.
5. 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: 1 μL in 100 μL 1X PBS containing 0.5% BSA (green).
6. Blank control: Mouse Kidney (blue). Primary Antibody: Rabbit Anti-phospho-Fas Ligand antibody (TMAB-00655,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 1 X PBS containing 0.5% BSA.
Protocol
The cells were fixed with 2% paraformaldehyde for 10 min at 37°C. Primary antibody (TMAB-00655, 1 μg/1x10^6 cells) were incubated for 30 min at room temperature, followed by 1 X PBS containing 0.5% BSA + 10% 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.
7. Sample: Bone (mouse) Lysate at 40 μg
Primary: Anti-Fas ligand (TMAB-00655) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 46 kDa
8. Sample:
Lane 1: Bone (Mouse) Lysate at 40 μg
Lane 2: Bone (Rat) Lysate at 40 μg
Primary: Anti-Fas Ligand (TMAB-00655) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDa
9. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (Fas Ligand) Polyclonal Antibody, Unconjugated (TMAB-00655) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human Fas Ligand

Human

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