|Product Name||Anti-P53 Monoclonal Antibody|
|Storage & Handling||At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.|
|Description||Mouse IgG monoclonal antibody for p53, tumor protein p53 (TP53) detection. Tested with WB, IHC-P, IHC-F, ICC in Human. No cross reactivity with other proteins.|
|Cite This Product||Anti-P53 Monoclonal Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # MA1078)|
|Contents/Buffer||Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as preservative.|
|Immunogen||Recombinant human wild-type p53 protein.|
Assay Dilutions Overview
Immunohistochemistry(Paraffin-embedded Section), 0.5-2μg/ml, Human, By Heat
Immunocytochemistry , 1μg/ml, Human, -
Immunohistochemistry(Frozen Section), 0.5-2μg/ml, Human, -
Western blot, 0.5-1μg/ml, Human
Boster's Secondary Antibodies And IHC, WB Kits
The following reagents are used to generate the images below.Boster recommends Enhanced Chemiluminescent Kit with anti-Mouse IgG (EK1001) for Western blot, and HRP Conjugated anti-Mouse IgG Super Vision Assay Kit (SV0001-1) for IHC(P), IHC(F) and ICC.
Images And Assay Conditions
Figure 1. Western blot analysis of p53 using anti-p53 antibody (MA1078).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: HL-60 whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-p53 antigen affinity purified monoclonal antibody (Catalog # MA1078) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for p53 at approximately 53KD. The expected band size for p53 is at 53KD.
Figure 2. IHC analysis of p53 using anti-p53 antibody (MA1078).
p53 was detected in paraffin-embedded section of human mammary cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-p53 Antibody (MA1078) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1021) with DAB as the chromogen.
Figure 3. IF analysis of p53 using anti- p53 antibody (MA1078)
p53 was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/mL mouse anti- p53 Antibody (MA1078) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Protein Target Info (Source: Uniprot.org)
|Protein Name||Cellular tumor antigen p53|
|Tissue Specificity||TP53: Ubiquitous. Isoforms are expressed in a widerange of normal tissues but in a tissue-dependent manner. Isoform2 is expressed in most normal tissues but is not detected inbrain, lung, prostate, muscle, fetal brain, spinal cord and fetalliver. Isoform 3 is expressed in most normal tissues but is notdetected in lung, spleen, testis, fetal brain, spinal cord andfetal liver. Isoform 7 is expressed in most normal tissues but isnot detected in prostate, uterus, skeletal muscle and breast.Isoform 8 is detected only in colon, bone marrow, testis, fetalbrain and intestine. Isoform 9 is expressed in most normal tissuesbut is not detected in brain, heart, lung, fetal liver, salivarygland, breast or intestine.|
|Alternative Names||Cellular tumor antigen p53;Tumor suppressor p53;Tp53;P53;|
|Subcellular Localization||Cytoplasm . Nucleus . Endoplasmic reticulum . Mitochondrion matrix . Interaction with BANP promotes nuclear localization. Translocates to mitochondria upon oxidative stress (By similarity). .|
|Molecular Weight||43451 MW|
*if product is indicated to react with multiple species, protein info is based on the human gene.
|Protein 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. Prevents CDK7 kinase activity when associated to CAK complex in response to DNA damage, thus stopping cell cycle progression. 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. Regulates the circadian clock by repressing CLOCK-ARNTL/BMAL1-mediated transcriptional activation of PER2. .|
|Research Areas||Apoptosis, Cell Biology, Intracellular, P53 Pathway
*You can search these to find other products in these research areas.
|Background||The p53 tumor antigen is found in increased amounts in a wide variety of transformed cells. The protein is also detectable in many actively proliferating, nontransformed cells, but it is undetectable or present at low levels in resting cells. This protein induces cell cycle arrest or apoptosis in response to sublethal or severe DNA damage, respectively, by differential transcription of target genes and through transcription-independent apoptotic functions. The p53 protein contains 393 amino acids. Human p53 tumour antigen is Locatedto band 17p13. p53 mutations are common in pancreatic cancer and are absent in chronic pancreatitis.|
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