Anti-A3AR Polyclonal Antibody is a Rabbit antibody targeting A3AR. Anti-A3AR Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Dog,Pig,Cow,Rabbit,Sheep)

1. Tissue/cell: human gastric 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-ADORA3 Polyclonal Antibody, Unconjugated (TMAB-00017) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (Adenosine A3 Receptor) Polyclonal Antibody, Unconjugated (TMAB-00017) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 min and DAB staining.
3. Sample:
Lane 1: Heart (Mouse) Tissue Lysate at 40 μg
Lane 2: Bone (Mouse) Tissue Lysate at 40 μg
Lane 3: Heart (Rat) Tissue Lysate at 40 μg
Primary:
Anti-Adenosine A3 Receptor (TMAB-00017) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 36 kDa
4. Blank control: HL-60. Primary Antibody (green line): Rabbit Anti-Adenosine A3 Receptor antibody (TMAB-00017)
Dilution: 2 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 1 μg/test.
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.
5. Blank control (blue line): Raji (blue)(The cells were fixed with 70% ethanol overnight at-20°C). Primary Antibody (green line): Rabbit Anti-Adenosine A3 Receptor antibody (TMAB-00017); Dilution: 1 μg/10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE;Dilution: 1 μg/test.

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 ADORA3

Human

More GPCR,Heart disease,Cardiac metabolism,Ischemia / Reperfusion