Name: Anti-CD45 antibody [EP322Y]
SKU: ab40763
Rating: 5 (14 reviews)

Anti-CD45 antibody [EP322Y] is a rabbit recombinant monoclonal antibody that is used to detect CD45 in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 50 publications
- Trusted since 2006

Images

Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] (AB40763), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

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

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/5000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/15 - 1/20	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Permeabilization and intracellular staining is necessary.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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Target data

See full target information PTPRC

Function

Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor (PubMed:35767951). 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

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

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
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

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-CD45 antibody [EP322Y] (ab40763) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Abcam's high quality manufacturing and validation processes ensure Anti-CD45 antibody [EP322Y] (ab40763) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-CD45 antibody [EP322Y] (ab40763) has 13 independent reviews from customers.

Anti-CD45 antibody [EP322Y] (ab40763) specifically detects CD45 (UniProt ID: P08575; Molecular weight: 145kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Conjugation-ready, carrier free format available for antibody clone EP322Y - Anti-CD45 antibody [EP322Y] - BSA and Azide free ab214437.

Antibody clone EP322Y is also available pre-conjugated to a variety of labels for your convenience - Alexa Fluor^® 488, Alexa Fluor^® 647, PE, PerCP, APC, Alexa Fluor^® 594 (Alexa Fluor® 488 Anti-CD45 antibody [EP322Y] ab200315, Alexa Fluor® 647 Anti-CD45 antibody [EP322Y] ab200317, PE Anti-CD45 antibody [EP322Y] ab214501, PerCP Anti-CD45 antibody [EP322Y] ab220298, APC Anti-CD45 antibody [EP322Y] ab221245, Alexa Fluor® 594 Anti-CD45 antibody [EP322Y] ab303598).

CD45 regulates immune cell signaling and is crucial in the development and function of immune cells, with alterations linked to various immunodeficiencies and autoimmune diseases.

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.

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.

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15 product images

Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] (ab40763)
Overlay histogram showing Jurkat (Human T cell leukemia cell line from peripheral blood) 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).
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)
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, 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 knockout cell line ab274932 (knockout cell lysate ab274990). The band observed in the knockout 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 knockout 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] (ab40763) at 1/5000 dilution
Lane 1: Wild-type Jurkat cell lysate at 20 µg
Lane 2: PTPRC knockout 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.
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)
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, 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] (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] (ab40763)
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 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] (ab40763)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labelling CD45 with 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)
Unpurified ab40763 showing negative staining in normal kidney tissue.
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)
Blocking/Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution
All lanes: HuT-78 cell lysate at 20 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 147 kDa
Observed band size: 200 kDa
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)
Blocking/Dilution buffer: 5% NFDM/TBST
All lanes: Western blot - Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution
All lanes: Raji (Human Burkitt's lymphoma cell line) cell lysate at 20 µg
Secondary
All lanes: HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 147 kDa
Observed band size: 200 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)
Unpurified ab40763 showing negative staining in skeletal muscle tissue.
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)
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 ECL Substrate Kit (High Sensitivity) ab133406.
All lanes: Western blot - Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution
All lanes: Jurkat (Human T cell lymphoblast-like cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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
Exposure time: 8min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)
Unpurified ab40763 showing positive staining in normal colon lymphoid cells tissue.
OI-RD Scanning - Anti-CD45 antibody [EP322Y] (ab40763)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)
Unpurified ab40763 showing positive staining in normal spleen tissue.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)
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 (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500. The sample is counterstained 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 (shown in the inset).
Flow Cytometry (Intracellular) - Anti-CD45 antibody [EP322Y] (ab40763)
Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab40763 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs 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 ab40763 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 10⁶ in 100 µl at 0.2 μg/ml (1/10,500)) for 30 min at 22°C .
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on live cells.