Overview

Description

  • Nature
    Recombinant
  • Source
    Baculovirus infected Sf9 cells
  • Amino Acid Sequence
    • Accession
    • Species
      Human
    • Molecular weight
      79 kDa including tags
    • Amino acids
      1 to 393
    • Tags
      GST tag N-Terminus

Specifications

Our Abpromise guarantee covers the use of ab84768 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

  • Applications

    Western blot

    SDS-PAGE

  • Form
    Liquid
  • Concentration information loading...

Preparation and Storage

  • Stability and Storage

    Shipped on dry ice. Upon delivery aliquot and store at -80ºC. Avoid freeze / thaw cycles.

    pH: 7.50
    Constituents: 0.307% Glutathione, 0.00174% PMSF, 0.00385% DTT, 0.79% Tris HCl, 0.00292% EDTA, 25% Glycerol, 0.29% Sodium chloride

General Info

  • Alternative names
    • Antigen NY-CO-13
    • BCC7
    • Cellular tumor antigen p53
    • FLJ92943
    • LFS1
    • Mutant tumor protein 53
    • p53
    • p53 tumor suppressor
    • P53_HUMAN
    • Phosphoprotein p53
    • Tp53
    • Transformation related protein 53
    • TRP53
    • Tumor protein 53
    • Tumor protein p53
    • Tumor suppressor p53
    see all
  • 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. Implicated in Notch signaling cross-over. 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.
  • Tissue specificity
    Ubiquitous. Isoforms are expressed in a wide range of normal tissues but in a tissue-dependent manner. Isoform 2 is expressed in most normal tissues but is not detected in brain, lung, prostate, muscle, fetal brain, spinal cord and fetal liver. Isoform 3 is expressed in most normal tissues but is not detected in lung, spleen, testis, fetal brain, spinal cord and fetal liver. Isoform 7 is expressed in most normal tissues but is not detected in prostate, uterus, skeletal muscle and breast. Isoform 8 is detected only in colon, bone marrow, testis, fetal brain and intestine. Isoform 9 is expressed in most normal tissues but is not detected in brain, heart, lung, fetal liver, salivary gland, breast or intestine.
  • Involvement in disease
    Note=TP53 is found in increased amounts in a wide variety of transformed cells. TP53 is frequently mutated or inactivated in about 60% of cancers. TP53 defects are found in Barrett metaplasia a condition in which the normally stratified squamous epithelium of the lower esophagus is replaced by a metaplastic columnar epithelium. The condition develops as a complication in approximately 10% of patients with chronic gastroesophageal reflux disease and predisposes to the development of esophageal adenocarcinoma.
    Defects in TP53 are a cause of esophageal cancer (ESCR) [MIM:133239].
    Defects in TP53 are a cause of Li-Fraumeni syndrome (LFS) [MIM:151623]. LFS is an autosomal dominant familial cancer syndrome that in its classic form is defined by the existence of a proband affected by a sarcoma before 45 years with a first degree relative affected by any tumor before 45 years and another first degree relative with any tumor before 45 years or a sarcoma at any age. Other clinical definitions for LFS have been proposed (PubMed:8118819 and PubMed:8718514) and called Li-Fraumeni like syndrome (LFL). In these families affected relatives develop a diverse set of malignancies at unusually early ages. Four types of cancers account for 80% of tumors occurring in TP53 germline mutation carriers: breast cancers, soft tissue and bone sarcomas, brain tumors (astrocytomas) and adrenocortical carcinomas. Less frequent tumors include choroid plexus carcinoma or papilloma before the age of 15, rhabdomyosarcoma before the age of 5, leukemia, Wilms tumor, malignant phyllodes tumor, colorectal and gastric cancers.
    Defects in TP53 are involved in head and neck squamous cell carcinomas (HNSCC) [MIM:275355]; also known as squamous cell carcinoma of the head and neck.
    Defects in TP53 are a cause of lung cancer (LNCR) [MIM:211980].
    Defects in TP53 are a cause of choroid plexus papilloma (CPLPA) [MIM:260500]. Choroid plexus papilloma is a slow-growing benign tumor of the choroid plexus that often invades the leptomeninges. In children it is usually in a lateral ventricle but in adults it is more often in the fourth ventricle. Hydrocephalus is common, either from obstruction or from tumor secretion of cerebrospinal fluid. If it undergoes malignant transformation it is called a choroid plexus carcinoma. Primary choroid plexus tumors are rare and usually occur in early childhood.
    Defects in TP53 are a cause of adrenocortical carcinoma (ADCC) [MIM:202300]. ADCC is a rare childhood tumor of the adrenal cortex. It occurs with increased frequency in patients with the Beckwith-Wiedemann syndrome and is a component tumor in Li-Fraumeni syndrome.
  • Sequence similarities
    Belongs to the p53 family.
  • Domain
    The nuclear export signal acts as a transcriptional repression domain. The TADI and TADII motifs (residues 17 to 25 and 48 to 56) correspond both to 9aaTAD motifs which are transactivation domains present in a large number of yeast and animal transcription factors.
  • Post-translational
    modifications
    Acetylated. Acetylation of Lys-382 by CREBBP enhances transcriptional activity. Deacetylation of Lys-382 by SIRT1 impairs its ability to induce proapoptotic program and modulate cell senescence.
    Phosphorylation on Ser residues mediates transcriptional activation. Phosphorylated by HIPK1 (By similarity). Phosphorylation at Ser-9 by HIPK4 increases repression activity on BIRC5 promoter. Phosphorylated on Thr-18 by VRK1. Phosphorylated on Ser-20 by CHEK2 in response to DNA damage, which prevents ubiquitination by MDM2. Phosphorylated on Thr-55 by TAF1, which promotes MDM2-mediated degradation. Phosphorylated on Ser-46 by HIPK2 upon UV irradiation. Phosphorylation on Ser-46 is required for acetylation by CREBBP. Phosphorylated on Ser-392 following UV but not gamma irradiation. Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylated on Ser-15 upon ultraviolet irradiation; which is enhanced by interaction with BANP.
    Dephosphorylated by PP2A-PPP2R5C holoenzyme at Thr-55. SV40 small T antigen inhibits the dephosphorylation by the AC form of PP2A.
    May be O-glycosylated in the C-terminal basic region. Studied in EB-1 cell line.
    Ubiquitinated by MDM2 and SYVN1, which leads to proteasomal degradation. Ubiquitinated by RFWD3, which works in cooperation with MDM2 and may catalyze the formation of short polyubiquitin chains on p53/TP53 that are not targeted to the proteasome. Ubiquitinated by MKRN1 at Lys-291 and Lys-292, which leads to proteasomal degradation. Deubiquitinated by USP10, leading to its stabilization. Ubiquitinated by TRIM24, which leads to proteasomal degradation. Ubiquitination by TOPORS induces degradation. Deubiquitination by USP7, leading to stabilization. Isoform 4 is monoubiquitinated in an MDM2-independent manner.
    Monomethylated at Lys-372 by SETD7, leading to stabilization and increased transcriptional activation. Monomethylated at Lys-370 by SMYD2, leading to decreased DNA-binding activity and subsequent transcriptional regulation activity. Lys-372 monomethylation prevents interaction with SMYD2 and subsequent monomethylation at Lys-370. Dimethylated at Lys-373 by EHMT1 and EHMT2. Monomethylated at Lys-382 by SETD8, promoting interaction with L3MBTL1 and leading to repress transcriptional activity. Demethylation of dimethylated Lys-370 by KDM1A prevents interaction with TP53BP1 and represses TP53-mediated transcriptional activation.
    Sumoylated by SUMO1.
  • Cellular localization
    Cytoplasm; Cytoplasm. Nucleus. Nucleus > PML body. Endoplasmic reticulum. Interaction with BANP promotes nuclear localization. Recruited into PML bodies together with CHEK2; Nucleus. Cytoplasm. Localized in both nucleus and cytoplasm in most cells. In some cells, forms foci in the nucleus that are different from nucleoli; Nucleus. Cytoplasm. Localized in the nucleus in most cells but found in the cytoplasm in some cells; Nucleus. Cytoplasm. Localized mainly in the nucleus with minor staining in the cytoplasm; Nucleus. Cytoplasm. Predominantly nuclear but localizes to the cytoplasm when expressed with isoform 4 and Nucleus. Cytoplasm. Predominantly nuclear but translocates to the cytoplasm following cell stress.
  • Information by UniProt

Images

  • SDS-PAGE showing ab84768 at approximately 79kDa.

References

ab84768 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Answer


You are correct, based on the molecular weight alone of the total amino acids in the protein it should be less than 79 kDa. Sometimes proteins can migrate a bit slower in the gel due to post translational modifications and the specific gel % and running conditions used, which in this case caused the protein to run around 79 kDa in SDS-PAGE and around 75-76 kDa in WB shown on the datasheet below:


https://www.abcam.com/recombinant-human-p53-protein-ab84768.html



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.



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Answer

Thank you for your email.

I am sorry I will not able to confirm the binding between ab80645 and ab84768 in ELISA; we unfortunately haven't tested these product in ELISA. These products however did show positive results in western blot.

I agree that ELISA is the quickest way to check the binding however it needs optimization. So I would suggest optimizing the dilutions of antibody and protein; the optimized concentration/ dilutionwill ultimately help in your required set of experiment.

I hope these suggestions would be helpful. Please do not hesitate to cotnact me if the result does not improve.

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Question

Hi,
please find the information

1. Order Details
Antibody code: ab80645
Problem
Choose: Problem with standard curve No signal or weak signal High background
Lot number
Purchase order number
or preferably Abcam order number
General Information
Antibody storage conditions (temperature/reconstitution etc) -80°C.
Description of the problem (high background, no signal, non-specific color development, poor standard curve etc.)
Type of ELISA (Direct ELISA/Indirect ELISA/Sandwich ELISA etc.) Indirect ELISA
Sample (Species/Cell type/Cell line etc.) p53 protein (Tagged) (ab84768)
Coating well (Buffer/Concentration of the coating material etc.) PBS pH=7.4
Blocking conditions (Buffer/time period, Blocking agent etc.) 1% BSA in PBS buffer
Primary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time, Wash step

Anti-p53 antibody [DO-1] - BSA and Azide free (ab80645), 1/1000, 1h,Secondary Antibody (Manufacturer/Species/Diluent/Dilution/Incubation time,

invitrogen, goat anti-mouse IgG1 (g1), horseradish peroxidase conjugate, PBS buffer pH=7.4, 1h
Detection method (Substrate/Diluent etc.)

TMB stabilized chromogen
Positive and negative controls used (please specify)
p53 protein and PBS
Optimization attempts (problem solving)
How many times have you tried the ELISA?

Once
Have you run a "No Primary" control?
Yes No
Do you obtain the same results every time?
Yes No
What steps have you altered?
dditional Notes:
Data:
We would appreciate if you are able to provide a copy of the data, particularly from the standard curve. This will help us to assess the results.
Actually, I did not measure the OD; I just compare the color change with the control by naked eye.

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Answer

Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody an protein.

I have indeed mentioned that these products are tested together, the application I meant was western blot. This information is present on the daasheet of both ab80645 and ab84768. I apologize if I was unclear. These two products are never been used in ELISA together so you will need to optimize the protocol. I have follow suggestions for protocol optimizations and hoping for best results;

- We recommend using coating buffer 8M Urea, 20mM Tris, 0.5M Nacl, pH 9.6 for coating the recombinant protein on the ELISA plate.
-Place the plate on rotating platform and incubate for 2 hours at RT or overnight at 4C.
- Gently wash the plate with PBST buffer
- Block the plate with 3% BSA in PBST for 1-2 hours at RT
- Wash the plate 3 times and apply primary antibody at different dilutions e.g. 1/100, 1/500 and 1/1000; the range will helps to get optimized dilution. Incubate the plate 1-4 hours at RT or 4C.
- Wash the plate and add secondary antibody for 1 hour, try different dilutions also
- Wash the plate 3 times with PBST and add detection reagent as required.
- read the plate using ELISA reador

Please note the concentrations of both protein and antibody should be optimized.. Please try the range of dilutions for both protein and antibody.

I am sure these suggestions would help to improve the results. Please do not hesitate to contact me if both products fails again.

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Answer

Thank you for contacting us.

ab26 recognizes both mutant and wild type forms of human p53. We unfortunately haven't used ab26 together with ab84768 and ab73735, so we are unsure about the performance of these products together. Theoretically these products should work because the antibody ab26 is raised against the immunogen, which is compatible with the proteins.

We however have tested ab80645 and ab84768 together so we can guarantee that there products will work together. The image is present on the datasheet.

All of these products comes with full money back "guarantee" when used in applications as stated on the datasheet so you can purchase and use these without hesitation.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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