Anti-PIRH2 Polyclonal Antibody is a Rabbit antibody targeting PIRH2. Anti-PIRH2 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. Tissue/cell: human glioma tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-PIRH2 Polyclonal Antibody, Unconjugated (TMAB-11465) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
2. Tissue/cell: human lung carcinoma; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01 M, pH 6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37℃ for 20 min;
Incubation: Anti-PIRH2 Polyclonal Antibody, Unconjugated (TMAB-11465) 1: 200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining
3. Sample:
Lane 1: Mouse Stomach tissue lysates
Lane 2: Mouse Spleen tissue lysates
Primary: Anti-PIRH2 (TMAB-11465) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 30 kDa
Observed band size: 30 kDa
4. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (PIRH2) polyclonal Antibody, Unconjugated (TMAB-11465) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
5. Blank control (black line): Hela.
Primary Antibody (green line): Rabbit Anti-PIRH2 antibody (TMAB-11465)
Dilution: 2 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then 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. Acquisition of 20,000 events was performed.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100-500; IF: 1:100-500; FCM: 2μg/Test

Polyclonal

KLH conjugated synthetic peptide: human PIRH2

Human

RCHY1

RING finger and CHY zinc finger domain-containing protein 1

Mediates E3-dependent ubiquitination and proteasomal degradation of target proteins, including p53/TP53, HDAC1 and CDKN1B. Preferentially acts on tetrameric p53/TP53. Increases AR transcription factor activity. Contributes to the regulation of CDKN1B and p53/TP53 levels, and thereby contributes to the regulation of the cell cycle progression.