Recombinant Anti-CD45 antibody [EP322Y] (ab40763)
- Datasheet
- References (16)
- Protocols
Overview
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Product name
Anti-CD45 antibody [EP322Y]
See all CD45 primary antibodies -
Description
Rabbit monoclonal [EP322Y] to CD45 -
Host species
Rabbit -
Specificity
Anti-CD45 antibody [EP322Y] (ab40763) recognizes cytoplasmic domain of CD45.
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Tested applications
Suitable for: WB, Flow Cyt, ICC/IF, IHC-P, IHC-Frmore details -
Species reactivity
Reacts with: Human -
Immunogen
corresponding to Human CD45 aa 1250-1350.
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Positive control
- Flow Cyt : Jurkat cells. IHC-P: Human kidney, colon lymphoid, tonsil and spleen tissue. ICC/IF: Jurkat cells. WB: Jurkat, HuT-78 and Raji cell lysate.
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General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 3.60 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, PBS -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Monoclonal -
Clone number
EP322Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-CD45 antibody [EP322Y] (Alexa Fluor® 488) (ab200315)
- Anti-CD45 antibody [EP322Y] (Alexa Fluor® 647) (ab200317)
- Anti-CD45 antibody [EP322Y] - BSA and Azide free (ab214437)
- Anti-CD45 antibody [EP322Y] (Phycoerythrin) (ab214501)
- Anti-CD45 antibody [EP322Y] (PerCP) (ab220298)
- Anti-CD45 antibody [EP322Y] (Allophycocyanin) (ab221245)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab40763 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | 1/5000. Detects a band of approximately 200 kDa (predicted molecular weight: 147 kDa). | |
| Flow Cyt | 1/15 - 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. Permeabilization and intracellular staining is necessary. |
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| ICC/IF | 1/100. | |
| IHC-P | 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
| IHC-Fr | Use at an assay dependent concentration. |
Target
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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
modificationsHeavily N- and O-glycosylated. -
Cellular localization
Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts. - Information by UniProt
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Database links
- Entrez Gene: 5788 Human
- Omim: 151460 Human
- SwissProt: P08575 Human
- Unigene: 654514 Human
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Alternative names
- B220 antibody
- CD 45 antibody
- CD45 antibody
see all
Images
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![Flow Cytometry - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-253468-anti-cd45-antibody-ep322y-flow-cytometry.jpg)
Flow Cytometry - 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).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-253431-anti-cd45-antibody-ep322y-immunohistochemistry.jpg)
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 (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).
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![Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-253457-anti-cd45-antibody-ep322y-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD45 antibody [EP322Y] (ab40763)Immunofluorescence staining of Jurkat cells (Human T cell leukemia cell line from peripheral blood) with purified ab40763 at a working dilution of 1/100, counterstained with DAPI.
The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel.
The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100.
The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab40763 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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![Western blot - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-254393-anti-cd45-antibody-ep322y-western-blot.jpg)
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (purified) + HuT-78 cell lysate at 20 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 147 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-195141-anti-cd45-antibody-ep322y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)Unpurified ab40763 showing positive staining in normal colon lymphoid cells tissue.
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![Western blot - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-254392-anti-cd45-antibody-ep322y-western-blot.jpg)
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (purified) + Raji (Human Burkitt's lymphoma cell line) cell lysate at 20 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 147 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-195140-anti-cd45-antibody-ep322y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)Unpurified ab40763 showing positive staining in normal spleen tissue.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-195142-anti-cd45-antibody-ep322y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)Unpurified ab40763 showing negative staining in skeletal muscle tissue.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-195143-anti-cd45-antibody-ep322y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [EP322Y] (ab40763)Unpurified ab40763 showing negative staining in normal kidney tissue.
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![Western blot - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-232690-anti-cd45-antibody-ep322y-western-blot.jpg)
Western blot - Anti-CD45 antibody [EP322Y] (ab40763)Anti-CD45 antibody [EP322Y] (ab40763) at 1/5000 dilution (unpurified, blocked with 3% milk) + Jurkat (Human T cell lymphoblast-like cell line) whole cell lysate at 10 µg
Secondary
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 why is the actual band size different from the predicted?
Exposure time: 8 minutesThis 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 ab133406.
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![Other - Anti-CD45 antibody [EP322Y] (ab40763)](https://www.abcam.com/ps/products/40/ab40763/Images/ab40763-195144-anti-cd45-antibody-ep322y.jpg)
Other - Anti-CD45 antibody [EP322Y] (ab40763)Equilibrium disassociation constant (KD)
Learn more about KDGo here to learn more about KD:
https://www.abcam.com/index.html?pageconfig=resource&rid=15749
References
This product has been referenced in:
- Nicastro M et al. Fibrocytes in Chronic Periaortitis: A Novel Mechanism Linking Inflammation and Fibrosis. Arthritis Rheumatol 71:1913-1922 (2019). Read more (PubMed: 31233292) »
- Chen X et al. Comprehensive Gene Expression Analysis in NMIBC Using RNA-seq Reveals New Therapy Strategies. Front Oncol 9:523 (2019). Read more (PubMed: 31293967) »
