Antibody Type: Rabbit Polyclonal

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

Reactivity: Human,Mouse,Rabbit (predicted:Pig,Cow)

IgG

Human,Mouse,Rabbit (predicted:Pig,Cow)

1. Tissue/cell: human colon 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-GPCR EX33 Polyclonal Antibody, Unconjugated (TMAB-00800) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: human brain glioma; 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-EX33 Polyclonal Antibody, Unconjugated (TMAB-00800) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.

3. Tissue/cell: rabbit thymus 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-EX33 Polyclonal Antibody, Unconjugated (TMAB-00800) 1:500, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
4. Blank control (blue): Mouse thymus cells (fixed with 2% paraformaldehyde (10 min)). Primary Antibody: Rabbit Anti-GPCR EX33 antibody (TMAB-00800), Dilution: 1 μg in 100 uL 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.
5. Paraformaldehyde-fixed, paraffin embedded (Human Brain Glioma); 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 (GPCR EX33) Polyclonal Antibody, Unconjugated (TMAB-00800) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
6. araformaldehyde-fixed, paraffin embedded (rabbit thymus); 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 (GPCR EX33) Polyclonal Antibody, Unconjugated (TMAB-00800) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
7. Blank control: U937. Primary Antibody (green line): Rabbit Anti-GPR84 antibody (TMAB-00800)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.

IHC-P,IHC-Fr,IF,FCM

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

Polyclonal

KLH conjugated synthetic peptide: human GPCR EX33 protein

Human

53831

GPCR