Name: Anti-CD45 antibody [EPR20033]
SKU: ab208022
Rating: 5 (1 reviews)

Anti-CD45 antibody [EPR20033] (ab208022) is a rabbit monoclonal antibody that is used to detect CD45 in Western Blot, IP, IHC-P. Suitable for Human, Mouse samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivalled batch-batch consistency

-Over 10 publications

Images

Western blot - Anti-CD45 antibody [EPR20033] (AB208022), 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

	IP	WB	IHC-P
Human	Expected	Tested	Tested
Mouse	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

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

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. 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 (By similarity). Dephosphorylates LYN, and thereby modulates LYN activity.

Alternative names

CD45, Ly-5, Ptprc, Receptor-type tyrosine-protein phosphatase C, Leukocyte common antigen, Lymphocyte antigen 5, 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: EPR20033
Purification technique: Affinity purification Protein A
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

What is this antibody validated in?

Anti-CD45 antibody [EPR20033] (ab208022) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P) in Human, Mouse samples.

What is the molecular weight of CD45?

Anti-CD45 [EPR20033] (ab208022) specifically detects a band for CD45 (UniProt: P06800) at a molecular weight of 145kDa.

Trusted by the scientific community

Anti-CD45 [EPR20033] (ab208022) was first used in a scientific publication in 2016 and has been cited over 10 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Other related products

We have a range of other formats of antibody clone [EPR20033] also available for your convenience:
ab208022, Carrier free - ab229292, Alexa Fluor® 647 - ab305209

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

What are the advantages of a recombinant monoclonal antibody?
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.

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

10 product images

Western blot - Anti-CD45 antibody [EPR20033] (ab208022)
False colour image of Western blot: Anti-CD45 antibody [EPR20033] staining at 1/1000 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, ab208022 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 [EPR20033] (ab208022) at 1/1000 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EPR20033] (ab208022)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse spleen tissue sections labeling CD45 with purified ab208022 at 1/5000 dilution (0.14 ?g/ml). Heat mediated antigen retrieval was performed. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-CD45 antibody [EPR20033] (ab208022)
False colour image of Western blot: Anti-CD45 antibody [EPR20033] staining at 1/1000 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, ab208022 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 [EPR20033] (ab208022) at 1/1000 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 [EPR20033] (ab208022)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 30 seconds; Lane 2: 5 seconds.
All lanes: Western blot - Anti-CD45 antibody [EPR20033] (ab208022) at 1/1000 dilution
Lane 1: J774A.1 (Mouse reticulum cell sarcoma macrophage cell line) whole cell lysate at 20 µg
Lane 2: WEHI-231 (Mouse B cell lymphoblast cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 147 kDa
Observed band size: 180 kDa
Immunoprecipitation - Anti-CD45 antibody [EPR20033] (ab208022)
CD45 was immunoprecipitated from 0.35 mg of J774A.1 (Mouse reticulum cell sarcoma macrophage cell line) whole cell lysate with ab208022 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab208022 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/1000 dilution.
Lane 1: J774A.1 whole cell lysate 10μg (Input).
Lane 2: ab208022 IP in J774A.1 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab208022 in J774A.1 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
All lanes: Immunoprecipitation - Anti-CD45 antibody [EPR20033] (ab208022)
Predicted band size: 147 kDa
Western blot - Anti-CD45 antibody [EPR20033] (ab208022)
Blocking/Dilution buffer: 5% NFDM/TBST.
293T and MCF7 are reported to be CD45 negative cell lines (PMID: 22978632& 24602188).
All lanes: Western blot - Anti-CD45 antibody [EPR20033] (ab208022) at 1/1000 dilution
Lane 1: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 2: 293T (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 3: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 147 kDa
Observed band size: 240 kDa
Exposure time: 3min
Western blot - Anti-CD45 antibody [EPR20033] (ab208022)
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2: 15 seconds.
All lanes: Western blot - Anti-CD45 antibody [EPR20033] (ab208022) at 1/1000 dilution
Lane 1: Mouse thymus lysate at 10 µg
Lane 2: Mouse spleen tissue lysate at 10 µg
Secondary
Lane 1: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Lane 2: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 147 kDa
Observed band size: 180 kDa, 240 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EPR20033] (ab208022)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD45 with ab208022 at 1/5000 (0.14 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human tonsil tissue.

The section was incubated with ab208022 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EPR20033] (ab208022)
Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CD45 with ab208022 at 1/5000 (0.14 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining in human kidney tissue.

The section was incubated with ab208022 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EPR20033] (ab208022)
Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling CD45 with ab208022 at 1/5000 (0.14 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: no staining in human skeletal muscle tissue.

The section was incubated with ab208022 for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.