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    cd45-antibody-i323-ab25386.pdf

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Immunology Adaptive Immunity B Cells CD
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Anti-CD45 antibody [I3/2.3] (ab25386)

  • Datasheet
  • SDS
Reviews (4)Q&A (4)References (37)

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

    Key features and details

    • Rat monoclonal [I3/2.3] to CD45
    • Suitable for: Flow Cyt
    • Reacts with: Mouse
    • Isotype: IgG2b

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    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: Flow Cytmore details
    • Species reactivity

      Reacts with: Mouse
    • Immunogen

      Tissue, cells or virus corresponding to CD45. T1M1 (Thy-1─c) cells

    • 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.

      The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    • Form

      Liquid
    • Storage instructions

      Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
    • Storage buffer

      pH: 8.20
      Constituent: 100% Borate buffered saline
    • Concentration information loading...
    • Purity

      Tissue culture supernatant
    • Clonality

      Monoclonal
    • Clone number

      I3/2.3
    • Isotype

      IgG2b
    • Research areas

      • Immunology
      • Adaptive Immunity
      • B Cells
      • CD
      • Immunology
      • Adaptive Immunity
      • T Cells
      • CD
      • Immunology
      • Cell Type Markers
      • CD
      • Non-lineage
      • Signal Transduction
      • Protein Phosphorylation
      • Tyrosine Phosphatases
      • Neuroscience
      • Cell Type Marker
      • Glia marker
      • Microglia marker
      • Stem Cells
      • Hematopoietic Progenitors
      • Surface Molecules
      • Stem Cells
      • Hematopoietic Progenitors
      • Lymphoid
      • B Lymphocytic Lineage
      • Stem Cells
      • Hematopoietic Progenitors
      • Lymphoid
      • T Lymphocytic Lineage
      • Stem Cells
      • Hematopoietic Progenitors
      • Lymphoid
      • NK Cell Lineage
      • Stem Cells
      • Hematopoietic Progenitors
      • Myeloid
      • Dendritic Cell Lineage
      • Stem Cells
      • Hematopoietic Progenitors
      • Myeloid
      • Monocytic Lineage
      • Stem Cells
      • Hematopoietic Progenitors
      • Myeloid
      • Neutrophil Lineage
      • Stem Cells
      • Hematopoietic Progenitors
      • Myeloid
      • Mast Cell Lineage
      • Immunology
      • Adaptive Immunity
      • Regulatory T Cells
      • Cardiovascular
      • Angiogenesis
      • Endothelial Cell Markers
      • Neuroscience
      • Development

    Associated products

    • Compatible Secondaries

      • Goat Anti-Rat IgG H&L (Alexa Fluor® 488) (ab150157)
      • Goat Anti-Rat IgG H&L (HRP) (ab205720)
    • Isotype control

      • Rat IgG2b, kappa monoclonal [RTK4530] - Isotype control (ab18541)
    • Recombinant Protein

      • Recombinant human CD45 protein (Fc Chimera Active) (ab214825)

    Applications

    The Abpromise guarantee

    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
    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.

    Notes
    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.
    • Target information above from: UniProt accession P08575 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 19264 Mouse
      • SwissProt: P06800 Mouse
      • Unigene: 391573 Mouse
      • 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

      • 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.

         

      Protocols

      • Flow cytometry protocols

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (37)

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

      ab25386 has been referenced in 37 publications.

      • Mori A  et al. Establishment of Human Leukocyte Antigen-Mismatched Immune Responses after Transplantation of Human Liver Bud in Humanized Mouse Models. Cells 10:N/A (2021). PubMed: 33672150
      • Pluchino N  et al. CXCR4 or CXCR7 antagonists treat endometriosis by reducing bone marrow cell trafficking. J Cell Mol Med 24:2464-2474 (2020). PubMed: 31904910
      • Martinez-Corral I  et al. Blockade of VEGF-C signaling inhibits lymphatic malformations driven by oncogenic PIK3CA mutation. Nat Commun 11:2869 (2020). PubMed: 32513927
      • Archacka K  et al. Beneficial Effect of IL-4 and SDF-1 on Myogenic Potential of Mouse and Human Adipose Tissue-Derived Stromal Cells. Cells 9:N/A (2020). PubMed: 32560483
      • Pérez-Hernández M  et al. Transcriptomic Coupling of PKP2 With Inflammatory and Immune Pathways Endogenous to Adult Cardiac Myocytes. Front Physiol 11:623190 (2020). PubMed: 33536940
      View all Publications for this product

      Customer reviews and Q&As

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

      1-8 of 8 Abreviews or Q&A

      Immunohistochemistry (Frozen sections) abreview for Anti-CD45 antibody [I3/2.3]

      Excellent
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Frozen sections)
      Sample
      Mouse Tissue sections (Colon (colitis))
      Permeabilization
      No
      Specification
      Colon (colitis)
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
      Fixative
      Acetone
      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.

      Abcam user community

      Verified customer

      Submitted Sep 17 2021

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

      Inconclusive
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Mouse Tissue sections (Bone marrow)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: Tested with citrate and Tris/EDTA
      Permeabilization
      No
      Specification
      Bone marrow
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
      Fixative
      Formaldehyde
      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.

      DR. Queralt Vallmajó

      Verified customer

      Submitted Mar 16 2020

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

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Mouse Tissue sections (Uterus)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: Sodium citrate
      Permeabilization
      Yes - Tween-20
      Specification
      Uterus
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
      Fixative
      Paraformaldehyde
      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.

      Reshef Tal

      Verified customer

      Submitted Sep 04 2019

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

      Excellent
      Abreviews
      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
      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.

      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

      Abcam community

      Verified customer

      Asked on Jan 14 2014

      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

      Jeremy Kasanov

      Abcam Scientific Support

      Answered on Jan 14 2014

      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

      Abcam community

      Verified customer

      Asked on May 01 2012

      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

      Abcam Scientific Support

      Answered on May 01 2012

      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

      Abcam community

      Verified customer

      Asked on Jan 31 2012

      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

      Read More

      Abcam Scientific Support

      Answered on Jan 31 2012

      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

      Abcam community

      Verified customer

      Asked on Oct 04 2011

      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

      Abcam Scientific Support

      Answered on Oct 04 2011

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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