p53 antibody (mAb)

RRID: AB_2793254
Clone: DO1
Aliases: TP53, BCC7, LFS1, TRP53
Catalog No: 39553 Format: 200 µg ¥70,000 Buy
Catalog No: 39554 Format: 10 µg ¥20,000 Buy

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Antibody Type:
Monoclonal
Isotype:
IgG
Purification:
Protein G Chromatography
Host:
Mouse
Molecular Weight:
53 kDa
Reactivity:
Human

Applications

ChIP Validated Western Blot Validated Immunoprecipitation Validated

Application Notes


Validated Applications:
IP: 5 µg per IP
WB: 0.5 - 2 µg/ml dilution

Published Applications:

ChIP

Also available as an AbFlex® engineered recombinant antibody.  For details on the corresponding AbFlex Recombinant Antibody, see Catalog No. 91247.

Immunogen

This p53 antibody was raised against full-length recombinant human p53 protein.

Buffer

Purified IgG in 70 mM Tris (pH 8), 105 mM NaCl, 31 mM glycine, 0.07 mM EDTA, 30% glycerol and 0.035% sodium azide. Sodium azide is highly toxic.

References

 

p53 mAb (Clone DO1) tested by Western blot.
5 µg of p53 mAb (Clone DO1) was used to immunoprecipitate p53 from 500 µg of camptothecin-treated U2OS nuclear extract (lane 2). 5 µg of mouse IgG was also used as a control (lane 1). The immunoprecipitated protein was detected by western blotting using the same p53 antibody.

p53 mAb (Clone DO1) tested by Western blot.
Western Blot: Nuclear extract of U2OS cells (20 μg) probed with p53 mAb (Clone DO1) (1 µg/ml).
    Lane 1: No treatment.
    Lane 2: Cells treated with camptothecin.

Background

p53 is the most important tumor suppressor in the genome. It is responsive to numerous genotoxic stresses, which activates its transcription factor activity, in turn causing cell-cycle arrest by activating expression of p21 Cip/WAF. Mutant p53 that has lost its DNA-binding function interferes with the activity of native p53 and leads to oncogenic transformation. Alternatively, transformation may be caused by overexpression of Mdm2/Hdm2, a ubiquitin ligase specific for p53, which causes its destabilization. Inactivation of p53 is often coincident with hyperactivation of NFκB (NFκB p50 and NFκB p65), both of which serve to inhibit apoptosis.

Positive Control

Nuclear extracts from U2OS cells treated with Camptothecin can be used as a positive control for p53 antibody (Clone DO1).

Chromatin IP Assays

This antibody has been validated for use in ChIP and/or ChIP-Seq, and can be used with Active Motif's ChIP-IT® High Sensitivity Kit or our magnetic bead-based ChIP-IT® Express Kits. For an overview of all of our ChIP products, please visit our Chromatin IP Page to learn about ChIP products designed for use with p53 antibody (mAb).

Bridging Antibody to Improve ChIP, MeDIP and IP

Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use p53 antibody (mAb).

Storage

Some products may be shipped at room temperature. This will not affect their stability or performance. Avoid repeated freeze/thaw cycles by aliquoting items into single-use fractions for storage at -20°C for up to 2 years. Keep all reagents on ice when not in storage.

Guarantee

This product is guaranteed for 12 months from date of receipt.

This product is for research use only and is not for use in diagnostic procedures.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • TR-FRET = Time-Resolved Fluorescence Resonance Energy Transfer;
  • WB = Western Blotting