Overview

  • Product nameAnti-CD45 antibody [EP322Y]
    See all CD45 primary antibodies
  • Description
    Rabbit monoclonal [EP322Y] to CD45
  • Specificityab40763 recognizes cytoplasmic domain of CD45.
  • Tested applicationsSuitable for: WB, Flow Cyt, ICC/IF, IHC-P, IHC-Frmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide corresponding to residues in the C-terminal domain of human CD45.

  • Positive control
    • Jurkat cells and cell lysate.
  • General notes

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab40763 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 1/5000. Detects a band of approximately 200 kDa (predicted molecular weight: 147 kDa).
Flow Cyt 1/15 - 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Permeabilization and intracellular staining is necessary.

ICC/IF 1/100.
IHC-P 1/250.
IHC-Fr Use at an assay dependent concentration.

Target

  • FunctionProtein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.
  • Involvement in diseaseDefects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
    Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200]. MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.
  • Sequence similaritiesBelongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 tyrosine-protein phosphatase domains.
  • DomainThe first PTPase domain interacts with SKAP1.
  • Post-translational
    modifications
    Heavily N- and O-glycosylated.
  • Cellular localizationMembrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt
  • Database links
  • Alternative names
    • B220 antibody
    • CD 45 antibody
    • CD45 antibody
    • CD45 antigen antibody
    • CD45R antibody
    • GP180 antibody
    • L-CA antibody
    • LCA antibody
    • Leukocyte common antigen antibody
    • loc antibody
    • Ly-5 antibody
    • LY5 antibody
    • Ly5, homolog of antibody
    • Lyt-4 antibody
    • OTTHUMP00000033813 antibody
    • OTTHUMP00000033816 antibody
    • OTTHUMP00000033817 antibody
    • OTTHUMP00000038574 antibody
    • Protein tyrosine phosphatase receptor type c polypeptide antibody
    • protein tyrosine phosphatase, receptor type, C antibody
    • Ptprc antibody
    • PTPRC_HUMAN antibody
    • Receptor-type tyrosine-protein phosphatase C antibody
    • T200 antibody
    • T200 glycoprotein antibody
    • T200 leukocyte common antigen antibody
    see all

Anti-CD45 antibody [EP322Y] images

  • Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (purified) + HuT-78 cell lysate at 20 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 147 kDa
    Observed band size : 200 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescence staining of Jurkat cells with purified ab40763 at a working dilution of 1/100, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab40763 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Immunohistochemical staining of paraffin embedded human tonsil with purified ab40763 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab40763 at a dilution of 1 in 20 (red line). The secondary antibody used was FITC goat anti-rabbit at a dilution of 1 in 500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  • Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (purified) + Raji cell lysate at 20 µg

    Secondary
    HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

    Predicted band size : 147 kDa
    Observed band size : 200 kDa (why is the actual band size different from the predicted?)

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Unpurified ab40763 showing positive staining in Normal spleen tissue.

  • Unpurified ab40763 showing positive staining in Normal colon lymphoid cells tissue.

  • Unpurified ab40763 showing negative staining in Skeletal muscle tissue.

  • Unpurified ab40763 showing negative staining in Normal kidney tissue.

  • Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (unpurified, blocked with 3% milk) + Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 147 kDa
    Observed band size : 200 kDa (why is the actual band size different from the predicted?)


    Exposure time : 8 minutes

    This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab40763 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Go here to learn more about KD:

    http://www.abcam.com/index.html?pageconfig=resource&rid=15749

     

References for Anti-CD45 antibody [EP322Y] (ab40763)

This product has been referenced in:
  • Chen YY  et al. Capture and Identification of Heterogeneous Circulating Tumor Cells Using Transparent Nanomaterials and Quantum Dots-Based Multiplexed Imaging. J Cancer 7:69-79 (2016). ICC/IF ; Human . Read more (PubMed: 26722362) »
  • Judd LM  et al. Elevated IL-33 expression is associated with pediatric eosinophilic esophagitis, and exogenous IL-33 promotes eosinophilic esophagitis development in mice. Am J Physiol Gastrointest Liver Physiol 310:G13-25 (2016). Mouse . Read more (PubMed: 26514775) »

See all 5 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (skin)
Permeabilization No
Specification skin
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative Formaldehyde
Username

Abcam user community

Verified customer

Submitted Aug 26 2016

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (human small intestine tissue)
Specification human small intestine tissue
Permeabilization No
Fixative Paraformaldehyde
Username

Dr. Irit Shoval

Verified customer

Submitted Sep 19 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Sample Human Tissue sections (Tonsil)
Specification Tonsil
Permeabilization No
Fixative Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

Submitted May 31 2014

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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DISCOUNT CODE: ***
Expiration date: November 24, 2012

This code will give you 1 free primary antibodybefore the expiration date. To redeem this offer, please submit an Abreview for ab40763 in ICC and include this code in the “Addit...

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