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Immunology Adaptive Immunity B Cells CD
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Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)

  • Datasheet
  • SDS
Submit a review Q&A (3)References (7)

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Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
  • Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
  • Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)

Key features and details

  • Mouse monoclonal [F10-89-4] to CD45 - Hematopoietic Stem Cell Marker
  • Suitable for: ICC/IF, Flow Cyt, IHC-P
  • Reacts with: Human
  • Isotype: IgG2a

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Overview

  • Product name

    Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker
    See all CD45 primary antibodies
  • Description

    Mouse monoclonal [F10-89-4] to CD45 - Hematopoietic Stem Cell Marker
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    See all applications and species data
  • Immunogen

    Tissue, cells or virus corresponding to Human CD45. Human T lymphocytes.

  • Positive control

    • IHC-P: Human tonsil (normal) tissue. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells, whole blood.
  • General notes

    Clone F10-89-4 reacts with all forms of CD45 expressed by all haematopoietic cells, except erythrocytes, having a higher level of expression on lymphocytes than on granulocytes.

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    IgG fraction
  • Primary antibody notes

    Clone F10-89-4 reacts with all forms of CD45 expressed by all haematopoietic cells, except erythrocytes, having a higher level of expression on lymphocytes than on granulocytes.
  • Clonality

    Monoclonal
  • Clone number

    F10-89-4
  • Myeloma

    NS1
  • Isotype

    IgG2a
  • 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

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-CD45 antibody [F10-89-4] (ab197730)
    • Anti-CD45 antibody [F10-89-4] - BSA and Azide free (ab230296)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG2a, Kappa Monoclonal [MOPC-173] - Isotype Control - ChIP Grade (ab18413)
    • Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18415)
  • Recombinant Protein

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab30470 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
Application Abreviews Notes
ICC/IF
Use a concentration of 5 - 10 µg/ml.
Flow Cyt
1/10 - 1/50.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-P
Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
ICC/IF
Use a concentration of 5 - 10 µg/ml.
Flow Cyt
1/10 - 1/50.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

IHC-P
Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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: 5788 Human
    • Omim: 151460 Human
    • SwissProt: P08575 Human
    • Unigene: 654514 Human
    • 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

    • Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
      Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)

      ab30470 staining CD45 in Jurkat (Human T cell leukemia cell line from peripheral blood) cells. The cells were fixed with 4% formaldehyde (10 minutes), then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1 hour. The cells were then incubated overnight at +4°C with ab30470 at 5 µg/ml (shown in green) and ab206369, Rabbit monoclonal to beta Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). This was followed by an incubation at room temperature for 1 hour with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed, at 1 µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

      This product also gave a positive signal under the same testing conditions in Jurkat cells fixed with 80% methanol (5 minutes).

    • Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
      Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)

      Flow cytometry staining of human whole blood with ab30470 (right) or mouse IgG2aκ; (ab18413) isotype (left). Red blood cells of 200 µL blood were lysed, then cells were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab30470) or mouse IgG2aκ; (ab18413) isotype (1x106 in 100 µL at 0.2 µg/ml) for 30 min on ice.

      The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor ® 488, pre-adsorbed) (ab150117) was used at 1/2000 dilution for 30 min on ice.

      Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter. Events were gated on alive cells.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)

      IHC image of CD45 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 minutes. The section was then incubated with ab30470, 1 µg/ml, for 15 minutes at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    • Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)
      Flow Cytometry - Anti-CD45 antibody [F10-89-4] - Hematopoietic Stem Cell Marker (ab30470)

      Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) cells stained with ab30470 (red line). The cells were fixed with 4% paraformaldehyde (10 minutes) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab30470, 1 µg/1 x 106 cells) for 30 minutes at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 minutes at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1 µg/1 x 106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 minutes) used under the same conditions.

      Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (7)

    Publishing research using ab30470? Please let us know so that we can cite the reference in this datasheet.

    ab30470 has been referenced in 7 publications.

    • Dong J  et al. Covalent chemistry on nanostructured substrates enables noninvasive quantification of gene rearrangements in circulating tumor cells. Sci Adv 5:eaav9186 (2019). PubMed: 31392269
    • Zhang F  et al. Defining inflammatory cell states in rheumatoid arthritis joint synovial tissues by integrating single-cell transcriptomics and mass cytometry. Nat Immunol 20:928-942 (2019). PubMed: 31061532
    • Desancé M  et al. Chondrogenic Differentiation of Defined Equine Mesenchymal Stem Cells Derived from Umbilical Cord Blood for Use in Cartilage Repair Therapy. Int J Mol Sci 19:N/A (2018). Flow Cyt ; Horse . PubMed: 29439436
    • Eriksson BO  et al. Upregulation of Plasminogen Activator Inhibitor-1 in Irradiated Recipient Arteries and Veins from Free Tissue Transfer Reconstruction in Cancer Patients. Mediators Inflamm 2018:4058986 (2018). PubMed: 30402041
    • Liu S  et al. Combined cell surface carbonic anhydrase 9 and CD147 antigens enable high-efficiency capture of circulating tumor cells in clear cell renal cell carcinoma patients. Oncotarget 7:59877-59891 (2016). PubMed: 27494883
    • Tharp WG  et al. Effects of glucose and insulin on secretion of amyloid-ß by human adipose tissue cells. Obesity (Silver Spring) 24:1471-9 (2016). PubMed: 27172814
    • Dalchau R  et al. Monoclonal antibody to a human leukocyte-specific membrane glycoprotein probably homologous to the leukocyte-common (L-C) antigen of the rat. Eur J Immunol 10:737-44 (1980). PubMed: 7428806

    Customer reviews and Q&As

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

    1-3 of 3 Abreviews or Q&A

    Question

    Ab30470 (anti CD45), does this recognise an intracellular or extracellular portion of the CD45 receptor?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 01 2017

    Answer

    The immunogen for ab30470 CD45 antibody [F10-89-4] is Tissue/ cell preparation of Human T lymphocytes. The epitope has not been mapped.

    The antibody is applicable for Flow cytometry. The protocol provided with the testing image on the datasheet indicates no permeabilization was required for the staining. Therefore, it is likely that it detects the extracellular portion of the CD45 receptor.

    Read More

    Abcam Scientific Support

    Answered on Aug 01 2017

    Question

    Customer contacted us regarding specific markers for white blood cells (ab30470) and cancer cells (ab7753). He observed CD45 staining by ab30470 in his cancer cells.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 20 2012

    Answer


    After performing an extensive search for leukocyte specific markers, there isn't 1 definitive marker that I can guarantee will only recognize leukocytes and not your cancer cells. With that being said, I have compiled a few options that I think will give you a better chance of having a marker that only recognizes your leukocytes and not your cancer cells based on the list of leukocyte markers given in the link below:
    http://www.antibodybeyond.com/reviews/cell-markers/leukocyte-marker.htm

    The antibodies that I compiled that I think may give you the best chance of finding a leukocyte-specific marker based on human protein atlas information are listed below:

    LAM-1 (SELL, CD62L): http://www.proteinatlas.org/ENSG00000188404/cell
    CD62L antibodies:
    1. https://www.abcam.com/cd62l-antibody-fmc46-ab11364.html
    2. https://www.abcam.com/cd62l-antibody-mel-14-ab119834.html

    CD18/CD11: http://www.proteinatlas.org/ENSG00000169896/cell
    1. https://www.abcam.com/cd11cd18-antibody-24-ab13219.html
    2. https://www.abcam.com/cd18-antibody-clb-lfa-11-54-ab51377.html

    I cannot guarantee that these antibodies will not label your cancer cells as well, but I hope the information is helpful to you and that the antibody beyond link listing leukocyte markers and the human protein atlas can help you in your search.

    Read More

    Abcam Scientific Support

    Answered on Dec 20 2012

    Question

    Customer contacted us regarding specific markers for white blood cells (ab30470) and cancer cells (ab7753). He observed CD45 staining by ab30470 in his cancer cells.

    Read More

    Abcam community

    Verified customer

    Asked on Dec 20 2012

    Answer


    Below is a link to the human protein atlas database file for CD45, I will forward the alternative specific marker information to you as soon as I compile it.
    http://www.proteinatlas.org/ENSG00000081237/cell

    Read More

    Abcam Scientific Support

    Answered on Dec 20 2012

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