p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95] - BSA and Azide free
Catalog# HA750920
p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95] - BSA and Azide free
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WB
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IF-Cell
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IP
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Mouse
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HA722074
含抗保成分
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unconjugated
Overview
Product Name
p53 Recombinant Rabbit Monoclonal Antibody [PSH03-95] - BSA and Azide free
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
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:1,000
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IF-Cell
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1:200
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IP
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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
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
ExpandImages
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☑ Cell treatment (CT)
Western blot analysis of p53 on different lysates with Rabbit anti-p53 antibody (HA750920) 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 (HA750920) 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. -
☑ Cell treatment (CT)
Immunocytochemistry analysis of NIH/3T3 cells treated with 0.5μM doxorubicin for 20 hours labeling p53 with Rabbit anti-p53 antibody (HA750920) 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 (HA750920) 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. -
☑ Cell treatment (CT)
p53 was immunoprecipitated in 0.2mg NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate with HA750920 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750920 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: HA750920 IP in NIH/3T3 treated with 0.5μM doxorubicincell for 24 hours cell lysate
Lane 3: Rabbit IgG instead of HA750920 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"