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p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95]

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Specification

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Catalog# HA722074

p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95]

Application

  • WB

  • IF-Cell

  • IP

Reactivity

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse p53 protein.

Species Reactivity

Mouse

Validated Applications

WB, IF-Cell, IP

Molecular Weight

Predicted band size: 53 kDa

Positive Control

RAW264.7 cell lysate, Neuro-2a cell lysate, NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours lysate, NIH/3T3.

Conjugation

unconjugated

Clone Number

PSH03-95

RRID

AB_3096714

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:200

  • IP

  • 1-2μg/sample

Target

Function

Acts as a tumor suppressor in many tumor types; induces growth arrest or apoptosis depending on the physiological circumstances and cell type. Involved in cell cycle regulation as a trans-activator that acts to negatively regulate cell division by controlling a set of genes required for this process. One of the activated genes is an inhibitor of cyclin-dependent kinases. Apoptosis induction seems to be mediated either by stimulation of BAX and FAS antigen expression, or by repression of Bcl-2 expression. In cooperation with mitochondrial PPIF is involved in activating oxidative stress-induced necrosis; the function is largely independent of transcription. Induces the transcription of long intergenic non-coding RNA p21 (lincRNA-p21) and lincRNA-Mkln1. LincRNA-p21 participates in TP53-dependent transcriptional repression leading to apoptosis and seem to have to effect on cell-cycle regulation. Implicated in Notch signaling cross-over. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. Isoform 2 enhances the transactivation activity of isoform 1 from some but not all TP53-inducible promoters. Isoform 4 suppresses transactivation activity and impairs growth suppression mediated by isoform 1. Isoform 7 inhibits isoform 1-mediated apoptosis. Regulates the circadian clock by repressing CLOCK-ARNTL/BMAL1-mediated transcriptional activation of PER2.

Background References

1. Louria-Hayon I et al. The promyelocytic leukemia protein protects p53 from Mdm2-mediated inhibition and degradation. J Biol Chem 278:33134-33141 (2003).

2. An W et al. Ordered cooperative functions of PRMT1, p300, and CARM1 in transcriptional activation by p53. Cell 117:735-748 (2004).

Subcellular Location

Nucleus. Cytoplasm. Localized in both nucleus and cytoplasm in most cells. In some cells, forms foci in the nucleus that are different from nucleoli.

UNIPROT

SwissProt: P02340 Mouse

Synonyms

Antigen NY-CO-13 antibody

BCC7 antibody

Cellular tumor antigen p53 antibody

FLJ92943 antibody

LFS1 antibody

Mutant tumor protein 53 antibody

p53 antibody

p53 tumor suppressor antibody

P53_HUMAN antibody

Phosphoprotein p53 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (<a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a>) at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: Neuro-2a cell lysate<br />Lane 3: NIH/3T3 cell lysate<br />Lane 4: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 53 kDa<br />Observed band size: 53 kDa<br /><br />Exposure time: 3 minutes 20 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (HA722074) at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: Neuro-2a cell lysate
    Lane 3: NIH/3T3 cell lysate
    Lane 4: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 53 kDa
    Observed band size: 53 kDa

    Exposure time: 3 minutes 20 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722074) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of NIH/3T3 cells treated with 0.5μM doxorubicin for 20 hours labeling p53 with Rabbit anti-p53 antibody (<a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p53 antibody (<a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a>) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of NIH/3T3 cells treated with 0.5μM doxorubicin for 20 hours labeling p53 with Rabbit anti-p53 antibody (HA722074) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-p53 antibody (HA722074) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />p53 was immunoprecipitated in 0.2mg NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate with <a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate (input)<br />Lane 2: <a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a> IP in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA722074" style="font-weight: bold;text-decoration: underline;">HA722074</a> in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 1 minute; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    ☑ Cell treatment (CT)

    p53 was immunoprecipitated in 0.2mg NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate with HA722074 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722074 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate (input)
    Lane 2: HA722074 IP in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate
    Lane 3: Rabbit IgG instead of HA722074 in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 1 minute; ECL: K1802

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation