Human Phospho-p53 (S46) Antibody

AF1489 1 Citation Datasheet / COA / SDS

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AF1489
AF1489-SP

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Human Phospho-p53 (S46) Antibody in Immunohistochemistry

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Phospho-p53 (S46) Antibody Summary

Species Reactivity

Human

Specificity

Detects human p53 when phosphorylated at S46.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

Phosphopeptide containing human p53 S46 site

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Simple Western

10 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Immunohistochemistry

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p53 in Human Breast Cancer Tissue.

p53 phosphorylated at S46 was detected in immersion fixed paraffin-embedded sections of human breast cancer tissue using Rabbit Anti-Human Phospho-p53 (S46) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1489) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Western Blot

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Detection of Human Phospho-p53 (S46) by Western Blot.

Western blot shows lysates of MCF-7 human breast cancer cell line untreated (-) or treated (+) with 1 µM camptothecin (CPT) for 5 hours. PVDF membrane was probed with 1 µg/mL Rabbit Anti-Human Phospho-p53 (S46) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1489) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for Phospho-p53 (S46) was detected at approximately 53 kDa (as indicated). The phospho-specificity of this antibody was supported by decreased labeling following treatment with 600 U lambda -phosphatase ( lambda -PPase) for 30 minutes. For additional reference the membrane was stripped and reprobed for total p53 using Human/Mouse/Rat p53 HRP-conjugated Antigen Affinity-purified Polyclonal Antibody (lower panel, Catalog # HAF1355). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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p53 in Human Breast.

p53 phosphorylated at S46 was detected in immersion fixed paraffin-embedded sections of human breast array using Rabbit Anti-Human Phospho-p53 (S46) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1489) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western

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Detection of Human Phospho-p53 (S46) by Simple Western^TM. Simple Western lane view shows lysates of MCF‑7 human breast cancer cell line untreated (-) or treated (+) with 1 µM Camptothecin (CPT) for 5 hours, loaded at 0.2 mg/mL. A specific band was detected for Phospho-p53 (S46) at approximately 65 kDa (as indicated) using 10 µg/mL of Rabbit Anti-Human Phospho-p53 (S46) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1489). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: p53

The p53 tumor suppressor protein signals apoptosis in cells that have incurred irreparable genotoxic damage. Phosphorylation of p53 at serine 46 is a critical event leading to the cellular decision to undergo apoptosis rather than cell cycle arrest. Serine 46 can be phosphorylated by either the p38 MAP kinase or the homeodomain-interacting protein kinase 2 (HIPK2).

Entrez Gene IDs

7157 (Human); 22059 (Mouse); 24842 (Rat)

Alternate Names

Antigen NY-CO-13; BCC7; FLJ92943; LFS1; LFS1TRP53; p53 tumor suppressor; p53; P53cellular tumor antigen p53; Phosphoprotein p53; TP53; transformation-related protein 53; TRP53; tumor protein p53; Tumor suppressor p53

Product Datasheets

Product Datasheet

COA

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Related Research Areas

Citation for Human Phospho-p53 (S46) Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

GRP-78 secreted by tumor cells blocks the antiangiogenic activity of bortezomib.
Authors: Kern J, Untergasser G, Zenzmaier C, Sarg B, Gastl G, Gunsilius E, Steurer M
Blood, 2009;114(18):3960-7.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot