Phospho-CDKN1A/p21 (Thr57) Rabbit Polyclonal Antibody

• Catalog Number: orb6569
• Category: Antibodies
• Description: Phospho-CDKN1A/p21 (Thr57) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated Synthesised phosphopeptide derived from human CDKN1A around the phosphorylation site of Thr57 TE(p-T)PL
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=2ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 18 kDa
• Target: CDKN1A
• UniProt ID: P38936
• RRID: AB_10922310
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: CDKN1A/p21(phospho Thr57); p21(phospho T57); p21(p
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Lehmann, Kuno et al. Liver failure after extended hepatectomy in mice is mediated by a p21-dependent barrier to liver regeneration Gastroenterology, 143, 1609-1619.e4 (2012)

PubMed: 22960658

Blank control: 293T. Primary Antibody (green line): Rabbit Anti-phospho-CDKN1A/p21 (Thr57) antibody (orb6569), 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 fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% PBST for 20 min at room temperature. 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 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (mouse lung), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-p21 (Thr57)) Polyclonal Antibody, Unconjugated (orb6569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse testis), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-p21 (Thr57)) Polyclonal Antibody, Unconjugated (orb6569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat lung), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-p21 (Thr57)) Polyclonal Antibody, Unconjugated (orb6569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat testis), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-p21 (Thr57)) Polyclonal Antibody, Unconjugated (orb6569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat uterus), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (phospho-p21 (Thr57)) Polyclonal Antibody, Unconjugated (orb6569) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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-phospho-CDKN1A/P21 (Thr57) Polyclonal Antibody, Unconjugated (orb6569) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.