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

IgG

Human,Mouse,Rat,Dog (predicted:Chicken,Pig,Cow,Horse)

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-PDGFRA Polyclonal Antibody, Unconjugated (TMAB-01354) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: rat colitis 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-PDGFRA Polyclonal Antibody, Unconjugated (TMAB-01354) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
3. Blank control: Mouse Kidney (blue). Primary Antibody: Rabbit Anti-PDGFRA antibody (TMAB-01354,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-01354, 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.
4. Black line: Positive blank control (SH-SY5Y); Negative blank control (A431)
Green line: Primary Antibody (Rabbit Anti-PDGFRA antibody (TMAB-01354))
Orange line: Isotype Control Antibody (Rabbit IgG).
Blue line: Secondary Antibody (Goat anti-rabbit IgG-AF488)
SH-SY5Y (Positive) and A431 (Negative control) cells (black) were fixed with 4% PFA for 10 min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at-20°C, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with PDGFRA Antibody (TMAB-01354) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody (blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

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 PDGF-R-A

Human

PDGF,Growth factor receptors,Growth Factors,Inflammatory mediators,PDGF,Receptor Tyrosine Kinases