Anti-CD45 antibody [EPR28934-536] 20 ul size (ab317446)

Rabbit Recombinant Monoclonal CD45 antibody. Suitable for ICC/IF, WB, Flow Cyt, IP, IHC-P and reacts with Mouse samples.

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Images

Western blot - Anti-CD45 antibody [EPR28934-536] (AB317446), expandable thumbnail

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

	ICC/IF	WB	Flow Cyt	IP	IHC-P
Human	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Mouse	Tested	Tested	Tested	Tested	Tested
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human, Rat	Dilution info -	Notes -

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, Receptor-type tyrosine-protein phosphatase C, Leukocyte common antigen, Lymphocyte antigen 5, T200, L-CA

Recommended products

Rabbit Recombinant Monoclonal CD45 antibody. Suitable for ICC/IF, WB, Flow Cyt, IP, IHC-P and reacts with Mouse samples.

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: EPR28934-536
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

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 [EPR28934-536] (ab317446)
CD45 Western blot staining using rabbit Anti-CD45 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: C2C12, HL-1.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 37917373).
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
All lanes: Western blot - Anti-CD45 antibody [EPR28934-536] (ab317446) at 1/1000 dilution
Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate at 80 µg with 5% NFDM/TBST
Lane 2: C2C12 (mouse myoblast) whole cell lysate at 80 µg with 5% NFDM/TBST
Lane 3: HL-1 (mouse atrial muscle cell) whole cell lysate at 80 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 200 kDa, 124 kDa
Exposure time: 180s
Western blot - Anti-CD45 antibody [EPR28934-536] (ab317446)
CD45 Western blot staining using rabbit Anti-CD45 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
Low expression: skeletal muscle.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 37917373).
In Western blot, Anti-Vinculin antibody [EPR8185] (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) staining at 1/10000 dilution.
All lanes: Western blot - Anti-CD45 antibody [EPR28934-536] (ab317446) at 1/1000 dilution
Lane 1: Mouse thymus tissue lysate at 80 µg with 5% NFDM/TBST
Lane 2: Mouse skeletal muscle tissue lysate at 80 µg with 5% NFDM/TBST
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Developed using the ECL technique.
Observed band size: 200 kDa, 124 kDa
Exposure time: 81s
Immunoprecipitation - Anti-CD45 antibody [EPR28934-536] (ab317446)
CD45 was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate with ab317446 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317446 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate

Lane 2: ab317446 IP in J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate

Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab317446 in J774A.1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CD45 antibody [EPR28934-536] (ab317446) at 1/30 dilution
All lanes: J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) whole cell lysate with 5% NFDM/TBST
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Exposure time: 180s
Flow Cytometry - Anti-CD45 antibody [EPR28934-536] (ab317446)
Flow cytometric analysis of Mouse PBMC cells labelling CD45 with ab317446 at 1/500 dilution (0.1ug) / right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti-CD3 conjugated to Alexa Fluor®647.

Gated on viable cells.
Flow Cytometry - Anti-CD45 antibody [EPR28934-536] (ab317446)
Flow cytometric analysis of J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) (Right) / HL-1 (mouse atrial muscle cell) (Left) cells labelling CD45 with ab317446 at 1/500 dilution (0.1ug)/Red 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).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Low expression: HL-1. Gated on vaible cells.
Flow Cytometry - Anti-CD45 antibody [EPR28934-536] (ab317446)
Flow cytometric analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (Right) / C2C12 (mouse myoblast) (Left) cells labelling CD45 with ab317446 at 1/500 dilution (0.1ug)/Red 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).
Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/5000 dilution was used as the secondary antibody.
Low expression: C2C12. Gated on viable cells.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EPR28934-536] (ab317446)
Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CD45 with ab317446 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Low expression tissue: Low expression on immune cells of mouse skeletal muscle.

The section was incubated with ab317446 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 [EPR28934-536] (ab317446)
Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD45 with ab317446 at 1/2000 (0.258 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on mouse spleen.

The section was incubated with ab317446 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
Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EPR28934-536] (ab317446)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse PBMC (mouse peripheral blood mononuclear cell) cells labelling CD45 with ab317446 at 1/500 (1.03 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green).
Confocal image showing membrane staining in mouse PBMC (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Anti-rat CD3 mouse monoclonal antibody (Alexa Fluor® 647) was used to counterstain tubulin at 1/100 5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EPR28934-536] (ab317446)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) cells labelling CD45 with ab317446 at 1/500 (1.03 ug/ml) dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/mL dilution (Green).
Confocal image showing membrane staining in J774A.1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).

Low expression: C2C12

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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 (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.