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  1. Link

    p53-antibody-epr17343-ab179477.pdf

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Cell Biology Cell Cycle Cell Cycle Inhibitors p53
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-p53 antibody [EPR17343] (ab179477)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (2) Submit a question References (28)

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Western blot - Anti-p53 antibody [EPR17343] (ab179477)
  • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR17343] (ab179477)
  • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
  • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
  • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
  • Anti-p53 antibody [EPR17343] (ab179477)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17343] to p53
  • Suitable for: WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Knockout
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Human TP53 (p53) knockout A549 cell line (ab276092)

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Overview

  • Product name

    Anti-p53 antibody [EPR17343]
    See all p53 primary antibodies
  • Description

    Rabbit monoclonal [EPR17343] to p53
  • Host species

    Rabbit
  • Specificity

    Based on the results of the knockout validation testing this antibody may not be suitable for IHC-P, Flow Cyt, IP or ChIP. Please contact our Scientific Support team for additional information.
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: A431, HEK-293, and T-47D whole cell lysates. ICC/IF: Wild-type HAP1 cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR17343
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Cell Cycle Inhibitors
    • p53
    • Cell Biology
    • Apoptosis
    • Intracellular
    • p53 Pathway
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA Damage & Repair
    • DNA Damage Response
    • p53
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Tumor Suppressors
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Cell Cycle Inhibitors
    • p53
    • Cancer
    • Cell cycle
    • Cell cycle inhibitors
    • p53 pathway
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • p53 pathway
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-p53 antibody [EPR17343] - BSA and Azide free (ab240215)
  • Assay kits

    • p53 Transcription Factor Assay Kit (Colorimetric) (ab207225)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant Human p53 protein (ab43615)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab179477 in the following tested applications.

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

Application Abreviews Notes
WB (2)
1/2000. Detects a band of approximately 53, 44 kDa (predicted molecular weight: 44 kDa).
ICC/IF
1/500.
Notes
WB
1/2000. Detects a band of approximately 53, 44 kDa (predicted molecular weight: 44 kDa).
ICC/IF
1/500.

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. 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.
  • Target information above from: UniProt accession P04637 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7157 Human
    • Omim: 191170 Human
    • SwissProt: P04637 Human
    • Unigene: 654481 Human
    • Alternative names

      • 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
      • Tp53 antibody
      • Transformation related protein 53 antibody
      • TRP53 antibody
      • tumor antigen p55 antibody
      • Tumor protein 53 antibody
      • Tumor protein p53 antibody
      • Tumor suppressor p53 antibody
      see all

    Images

    • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      Western blot - Anti-p53 antibody [EPR17343] (ab179477)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)

      Lane 2: p53 knockout HAP1 cell lysate (20 µg)

      Lane 3: A431 cell lysate (20 µg)

      Lane 4: Saos-2 cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab179477 observed at 53 kDa. Red - loading control, ab8226, observed at 42 kDa.

      ab179477 was shown to specifically react with p53 in wild type HAP1 cells along with additional cross reactive bands. No band was observed with p53 knockout samples were used.

      Wild-type and p53 knockout samples were subjected to SDS-PAGE. ab179477 and ab8226 (loading control to beta Actin) were diluted to 1/2000 and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR17343] (ab179477)
      Immunocytochemistry/ Immunofluorescence - Anti-p53 antibody [EPR17343] (ab179477)

      ab179477 staining p53 in wild-type HAP1 cells (top panel) and p53 knockout HAP1 cells (bottom panel). The cells were fixed with methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab179477 at 1/500 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      All lanes : Anti-p53 antibody [EPR17343] (ab179477) at 1/2000 dilution

      Lane 1 : Untreated HCT 116 (Human colorectal carcinoma) whole cell lysate
      Lane 2 : HCT 116 (Human colorectal carcinoma) whole cell lysate with 0.5µM Doxorubicin for 24hours whole cell lysate

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

      Predicted band size: 44 kDa
      Observed band size: 53 kDa why is the actual band size different from the predicted?

    • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      Anti-p53 antibody [EPR17343] (ab179477) at 1/20000 dilution + T-47D (Human ductal breast epithelial carcinoma cell line) whole cell lysates at 20 µg

      Secondary
      Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 44 kDa
      Observed band size: 44,53 kDa why is the actual band size different from the predicted?



      Blocking/dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      Western blot - Anti-p53 antibody [EPR17343] (ab179477)
      All lanes : Anti-p53 antibody [EPR17343] (ab179477) at 1/2000 dilution

      Lane 1 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysates
      Lane 2 : A431 (Human epidermoid carcinoma) whole cell lysates
      Lane 3 : Saos-2 (Human osteosarcoma cell line) whole cell lysates
      Lane 4 : HL-60 (Human promyelocytic leukemia cells) whole cell lysates
      Lane 5 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysates

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 44 kDa
      Observed band size: 53, 44 kDa why is the actual band size different from the predicted?



      Blocking and Diluting buffer and concentration: 5% NFDM/TBST.

      Saos-2, PC-3 and HL-60 cells are p53 null cell lines.

    • Anti-p53 antibody [EPR17343] (ab179477)
      Anti-p53 antibody [EPR17343] (ab179477)

    Protocols

    • Western blot protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (28)

    Publishing research using ab179477? Please let us know so that we can cite the reference in this datasheet.

    ab179477 has been referenced in 28 publications.

    • Cai T  et al. Notoginsenoside R1 induces DNA damage via PHF6 protein to inhibit cervical carcinoma cell proliferation. Mol Med Rep 23:N/A (2021). PubMed: 33537816
    • Li Z  et al. Juglone potentiates BRAF inhibitor-induced apoptosis in melanoma through reactive oxygen species and the p38-p53 pathway. Mol Med Rep 22:566-574 (2020). PubMed: 32377702
    • Hao Q  et al. Dual regulation of p53 by the ribosome maturation factor SBDS. Cell Death Dis 11:197 (2020). PubMed: 32198344
    • Su M  et al. Role of the p53-TRPM1/miR-211-MMP9 axis in UVB-induced human melanocyte migration and its potential in repigmentation. Int J Mol Med 45:1017-1026 (2020). PubMed: 31985026
    • Yu X  et al. TEOA Inhibits Proliferation and Induces DNA Damage of Diffuse Large B-Cell Lymphoma Cells Through Activation of the ROS-Dependent p38 MAPK Signaling Pathway. Front Pharmacol 11:554736 (2020). PubMed: 33013393
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-2 of 2 Abreviews or Q&A

    Western blot abreview for Anti-p53 antibody [EPR17343]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Colorectal cancer cell line)
    Gel Running Conditions
    Reduced Denaturing (12% gel)
    Loading amount
    30 µg
    Treatment
    10 µM CPT-11 for 24 hrs
    Specification
    Colorectal cancer cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Anke Rauch

    Verified customer

    Submitted Aug 24 2016

    Western blot abreview for Anti-p53 antibody [EPR17343]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (13%)
    Sample
    Human Cell lysate - whole cell (HCT116 human colon carcinoma cell line)
    Specification
    HCT116 human colon carcinoma cell line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MR. Christian Marx

    Verified customer

    Submitted Nov 19 2014

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