Skip to main content

Rabbit Monoclonal CD45 antibody. Carrier free. Suitable for ICC/IF, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 6 publications.


Images

Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] - BSA and Azide free (AB214437), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (AB214437), expandable thumbnail
  • Western blot - Anti-CD45 antibody [EP322Y] - BSA and Azide free (AB214437), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (AB214437), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] - BSA and Azide free (AB214437), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Constituents: PBS

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
ICC/IFWBFlow Cyt (Intra)IHC-P
Human
Tested
Tested
Tested
Tested

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species

Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info

-

Notes

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species

Human

Dilution info

-

Notes

Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

15 products for Alternative Product

6 products for Alternative Version

Target data

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. Dephosphorylates LYN, and thereby modulates LYN activity (By similarity).(Microbial infection) Acts as a receptor for human cytomegalovirus protein UL11 and mediates binding of UL11 to T-cells, leading to reduced induction of tyrosine phosphorylation of multiple signaling proteins upon T-cell receptor stimulation and impaired T-cell proliferation.

Alternative names

Receptor-type tyrosine-protein phosphatase C, Leukocyte common antigen, T200, L-CA, PTPRC, CD45

Recommended products

Rabbit Monoclonal CD45 antibody. Carrier free. Suitable for ICC/IF, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples. Cited in 6 publications.

Alternative names

Receptor-type tyrosine-protein phosphatase C, Leukocyte common antigen, T200, L-CA, PTPRC, CD45

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Carrier free

Yes

Clone number

EP322Y

Purification technique

Affinity purification Protein A

Specificity

This antibody recognizes cytoplasmic domain of CD45.

Dissociation constant

3.6 x 10-11 M

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

Notes

ab214437 is the carrier-free version of Anti-CD45 antibody [EP322Y] ab40763.

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

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

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD45 also known as Protein tyrosine phosphatase receptor type C (PTPRC) is a transmembrane glycoprotein with a molecular mass ranging between 180-240 kDa depending on its isoform. It is expressed on the surface of almost all hematopoietic cells except for mature erythrocytes and platelets. CD45 has a critical role in regulating antigen receptor signaling by modifying kinases involved in signal transduction and it is essential in lymphocyte development and activation. Because of its broad expression on immune cells CD45 is a valuable marker for differentiating various immune cell types in assays like flow cytometry and immunohistochemistry (IHC) often referred to as CD45 stain.

Biological function summary

CD45 acts by regulating tyrosine phosphorylation in the immune cell signaling context participating directly in signal transduction. It functions by dephosphorylating specific phosphotyrosine residues on various proteins modulating the signaling threshold required for lymphocyte activation. Although not known to be part of a protein complex CD45 itself shows isoform variation that associates with specific immune cell types impacting their function. Importantly CD45 interacts with multiple signaling molecules to affect cell growth and differentiation.

Pathways

Scientists associate CD45 with signaling pathways such as the T-cell receptor (TCR) and B-cell receptor (BCR) signaling. CD45 modulates the activity of Src family kinases important elements in these pathways making it essential for effective immune response and tolerance. Furthermore interactions between CD45 and proteins like Lck in T-cells or Lyn in B-cells highlight its pivotal role in executing its signaling functions.

Associated diseases and disorders

Scientists often link CD45 to immune-related conditions including autoimmunity and leukemia. In autoimmune diseases altered CD45 expression or activity can disrupt normal immune function contributing to pathogenesis. In leukemia CD45 expression levels can assist in disease classification and prognosis as it interacts with proteins involved in cell cycle regulation. The anti-CD45 antibodies can provide diagnostic and therapeutic avenues highlighting their utility in disease management.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

  • Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Clone EP322Y (ab214437) has been successfully conjugated by Abcam. This image was generated using Anti-CD45 antibody [EP322Y] (Alexa Fluor® 488). Please refer to Alexa Fluor® 488 Anti-CD45 antibody [EP322Y] ab200315 for protocol details.

    Alexa Fluor® 488 Anti-CD45 antibody [EP322Y] ab200315 staining CD45 in Daudi cells. The cells were fixed with 80% methanol (5 min), 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 overnight at +4°C with Alexa Fluor® 488 Anti-CD45 antibody [EP322Y] ab200315 at 1/250 dilution (shown in green) and Alexa Fluor® 647 Anti-Tubulin antibody [YOL1/34] - Microtubule Marker ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labeled 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 Daudi cells fixed with 4% formaldehyde (10 min).

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Clone EP322Y (ab214437) has been successfully conjugated by Abcam. This image was generated using Anti-CD45 antibody [EP322Y] (Alexa Fluor® 647). Please refer to Alexa Fluor® 647 Anti-CD45 antibody [EP322Y] ab200317 for protocol details.

    Overlay histogram showing Jurkat (human T cell leukemia cell line from peripheral blood) cells stained with Alexa Fluor® 647 Anti-CD45 antibody [EP322Y] ab200317 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (Alexa Fluor® 647 Anti-CD45 antibody [EP322Y] ab200317, 1/50000 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was Rabbit IgG (monoclonal) Alexa Fluor® 647 (Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab199093) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 670/14 bandpass filter.

    This antibody gave a positive signal in Jurkat cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

  • Western blot - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Western blot - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    False colour image of Western blot: Anti-CD45 antibody [EP322Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD45 antibody [EP322Y] ab40763 was shown to bind specifically to CD45. A band was observed at 160-220 kDa in wild-type Jurkat cell lysates with no signal observed at this size in PTPRC CRISPR-Cas9 edited cell line ab274932 (CRISPR-Cas9 edited cell lysate ab274990). The band observed in the CRISPR-Cas9 edited lysate lane below 160-220 kDa is likely to represent a truncated form of CD45. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and PTPRC CRISPR-Cas9 edited Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

    All lanes: Western blot - Anti-CD45 antibody [EP322Y] (Anti-CD45 antibody [EP322Y] ab40763) at 1/5000 dilution

    Lane 1: Wild-type Jurkat cell lysate at 20 µg

    Lane 2: PTPRC CRISPR-Cas9 edited Jurkat cell lysate at 20 µg

    Lane 3: THP-1 cell lysate at 20 µg

    Lane 4: MCF7 cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 147 kDa

    Observed band size: 160-220 kDa

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    This data was developed using Anti-CD45 antibody [EP322Y] ab40763, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell, Left) / Jurkat (Human T cell leukemia T lymphocyte, Right) cells labelling CD45 with Anti-CD45 antibody [EP322Y] ab40763 at 1/500 dilution (0.1ug)/ Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: 293T. (PMID: 16005866)

  • Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    This data was developed using Anti-CD45 antibody [EP322Y] ab40763, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labelling CD45 with Anti-CD45 antibody [EP322Y] ab40763 at 1/100 (6.1 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 2ug/ml dilution (Green). Confocal image showing membranouse staining in Jurkat cells and no staining in 293T cells. Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1000 2ug/ml dilution.

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified Anti-CD45 antibody [EP322Y] 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). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD45 antibody [EP322Y] ab40763).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Unpurified Anti-CD45 antibody [EP322Y] ab40763 showing negative staining in Normal kidney tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD45 antibody [EP322Y] ab40763).

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Clone EP322Y (ab214437) has been successfully conjugated by Abcam. This image was generated using Anti-CD45 antibody [EP322Y] (PE). Please refer to PE Anti-CD45 antibody [EP322Y] ab214501 for protocol details.

    Overlay histogram showing Jurkat (human T cell leukemia cell line from peripheral blood) cells stained with PE Anti-CD45 antibody [EP322Y] ab214501 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (PE Anti-CD45 antibody [EP322Y] ab214501, 1/500 dilution) for 30 min at 22°C.

    Isotype control antibody (black line) was Rabbit IgG (monoclonal) Phycoerythrin (PE Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab209478) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50 mW Yellow/Green laser (561nm) and 586/15 bandpass filter.

    This antibody gave a positive signal in Jurkat cells fixed with 4% formaldehyde (10 min)/permeabilized with 0.1% PBS-Triton X-100 for 15 min used under the same conditions.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Unpurified Anti-CD45 antibody [EP322Y] ab40763 showing negative staining in Skeletal muscle tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD45 antibody [EP322Y] ab40763).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Unpurified Anti-CD45 antibody [EP322Y] ab40763 showing positive staining in Normal colon lymphoid tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD45 antibody [EP322Y] ab40763).

  • OI-RD Scanning - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    OI-RD Scanning - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Equilibrium disassociation constant (KD)
    Learn more about KD

    Go here to learn more about KD:

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD45 antibody [EP322Y] ab40763).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    Unpurified Anti-CD45 antibody [EP322Y] ab40763 showing positive staining in Normal spleen tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD45 antibody [EP322Y] ab40763).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)

    This IHC data was generated using the same anti-CD45 antibody clone, EP322Y, in a different buffer formulation (cat# Anti-CD45 antibody [EP322Y] ab40763).

    Immunohistochemical staining of paraffin embedded human tonsil with purified Anti-CD45 antibody [EP322Y] ab40763 at a working dilution of 1/250. The secondary antibody used is HRP goat anti-rabbit IgG H&L (Goat Anti-Rabbit IgG H&L (HRP) 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.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com