Anti-CD45 antibody [I3/2.3] (ab25386)

Rat monoclonal CD45 antibody [I3/2.3]. Validated in IP, IHC, Flow Cyt and tested in Mouse. Cited in 17 publication(s). Independently reviewed in 1 review(s).

Reviews (1)Q&A (4)References (22)

Overview

  • Product name
    Anti-CD45 antibody [I3/2.3]
    See all CD45 primary antibodies
  • Description
    Rat monoclonal [I3/2.3] to CD45
  • Host species
    Rat
  • Tested applications
    Suitable for: IP, IHC-Fr, IHC-P, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Epitope
    ab25386 recognizes a framework epitope present on all CD45 isoforms.
  • Positive control
    • BALB/c spleen cells.
  • General notes

    This product was changed from ascites to tissue culture supernatant on (28 March 2019). Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Applications

Our Abpromise guarantee covers the use of ab25386 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
IP Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration. PubMed: 17579051
IHC-P Use at an assay dependent concentration. PubMed: 19695260
Flow Cyt Use at an assay dependent concentration.

See Haidl et al.

ab18536 - Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.

Target

  • Function
    Protein 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 disease
    Defects 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 similarities
    Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 tyrosine-protein phosphatase domains.
  • Domain
    The first PTPase domain interacts with SKAP1.
  • Post-translational
    modifications
    Heavily N- and O-glycosylated.
  • Cellular localization
    Membrane. 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
      • protein tyrosine phosphatase, receptor type, C antibody
      • Protein tyrosine phosphatase, receptor type, c polypeptide 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

    Images

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [I3/2.3] (ab25386)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [I3/2.3] (ab25386)Wong et al., PLoS, 9(6), e100770, fig6 ,doi: 10.1371/journal.pone.0100770

      ab25386 staining CD45 in E. coli infected Mouse prostate tissue sections 7 days post-instillation by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was blocked with 10% serum and 1% BSA for 1 hour at room temperature; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50 in blocking buffer) overnight at 4°C. An Alexa 488 conjugated Donkey polyclonal (1/100) was used as the secondary antibody. CD45 is stained green, Vimentin using ab92547 stained red, Hoechst nuclear stained blue. C is a high magnification of inset box in panel B.

      This image was generated using the ascites version of the product.

       

    • Flow Cytometry - Anti-CD45 antibody [I3/2.3] (ab25386)
      Flow Cytometry - Anti-CD45 antibody [I3/2.3] (ab25386)

      Flow Cytometry analysis of BALB/c mouse splenocytes labeling CD45 with ab25386 at 1 μg/106 cells (purple). A Mouse Anti-Rat IgG2b-PE was used as the secondary antibody. Grey - Isotype Control, Rat IgG2b-UNLB, followed by Mouse Anti-Rat IgG2b-PE.

      This image was generated using the ascites version of the product.

       

    References

    This product has been referenced in:
    • Poupeau A  et al. Genes controlling the activation of natural killer lymphocytes are epigenetically remodeled in intestinal cells from germ-free mice. FASEB J 33:2719-2731 (2019). Read more (PubMed: 30303739) »
    • Cazzamalli S  et al. Enhanced Therapeutic Activity of Non-Internalizing Small-Molecule-Drug Conjugates Targeting Carbonic Anhydrase IX in Combination with Targeted Interleukin-2. Clin Cancer Res 24:3656-3667 (2018). Read more (PubMed: 29691298) »
    See all 22 Publications for this product

    Publishing research using ab25386? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-5 of 5 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD45 antibody [I3/2.3]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate pH6
    Permeabilization
    No
    Specification
    tonsil
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    10% NBF
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 16 2017

    Question

    Based on the immunogen sequence, do you feel this antibody will likely cross react with human protein?

    Read More
    Answer

    The immunogen used to produce ab25386 was murine T1M1 (Thy-1-C) cells. This product is not routinely tested for cross reactivity with human; however, this clone was previously confirmed to be negative on human blood when tested by flow cytometry.

    Read More

    Question

    I am interested in purchasing ab25386 and was hoping to get more information regarding its use in IHC-P and Western blotting. The datasheet has no mention of WB -- is this because it was tested and failed or was it never tested? Also, it wasn't clear from the datasheet whether Abcam specifically tested IHC-P or if it was only listed based on its use in referenced papers. In other words, does the Abpromise guarantee cover IHC-P with this product?

    Read More
    Answer

    Thank you for contacting Abcam and for your interest in ab25386.


    Regarding WB, the antibody has not been tested in this application and therefore it is not covered by the Abpromise for WB. IHC-P has been added to the datasheet specifically due to the referenced publication. Although it has not been tested in house, we do guarantee the antibody for this application under the Abpromise.


    I hope this information is helpful. Please do not hesitate to contact us if you have any additional questions.

    Read More

    Question

    unsere Proben werden in 5-10% gepufferter Formalinlösung über Nacht fixiert und in der Pathologie eingebettet.
    Der Antikörper, der funktioniert ist anti CD 45 (rabbit) Nr. ab10558, das protokoll ist das gleiche,
    weil ich alle Antikörper in einem System brauche.
    Viele Grüße

    Read More
    Answer

    Vielen Dank für diese zusätzlichen Informationen.
    Ich würde Sie bitten die folgenden drei Tipps auszuprobieren und möchte Ihnen nochmals versichern, dass wenn die Protokolltipps das Resultat nicht verbessern, die Abpromise Garantie greift und ich Ihnen alternative Antikörper oder eine Rückerstattung anbieten kann.
    1.) Wir möchten vorschlagen einen "time course" für die Antigendemaskierung zu machen. Je nach Länge der Fixierungszeit, muss die Demaskierung angepasst werden. Leider sind bei der Antigendemaskierung keinerlei Regeln bekannt und es muss ausprobiert werden.
    2.) Da es sich bei allen Ihren Zielproteinen um Membranproteine handelt, empfehlen wir kein Permeabilisierungsreagenz zu verwenden. TWEEN wäscht Lipide aus der Membran aus und das könnte die Konfirmation der Markerproteine verändern.
    3.) Wir möchten auch empfehlen, 0.2M Glycin im Blockierungspuffer zu verwenden. Damit kann der Hintergrund, den Sie in der Milz sahen, vielleicht verringert werden. Glycin legt sich auf die freien Aldehydgruppen.
    Bitte lassen Sie mich wissen, ob diese Tipps geholfen haben oder nicht. Ich wünsche Ihnen viel Erfolg und verbleibe

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    Question

    To whom it may concern: I'm fixing in paraformaldehyde for 10 minutes after I scrape the cells off the cell culture plates. I do all staining after that. However, I don't think that the fixing step is the issue because my other two fluors are OK. I've attached a picture of my issue. The blue curve is the secondary control and the red is the stain of the positive control. My issue is that there isn't enough separation between the secondary control and positive control stain intensity. I think I used a farily high concentration of the CD45 primary antibody. Thanks for your help--it is much appreciated!

    Read More
    Answer

    Thanks for the additional information. At this time we have not had experience testing this antibody on rat samples. It has only been tested for reactivity to mouse CD45. Have you tested it on a mouse sample for a positive control?

    Read More

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