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CD45 Antibody – Mouse

For Research Use Only. Not for use in diagnostic procedures. Catalog #: MBS570368

The CD45 molecule is typically expressed at high levels on all hematopoietic cells. CD45 is a major component of the glycocalix of these cells and can be expressed in different isoforms. Antibody VIT200 recognizes a pan CD45 epitope, which is expressed on all… More details

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Product Overview

Online Datasheet in PDF
Product Name
CD45, Antibody
Full Product Name
Mouse anti CD45
Product Gene Name
Isotype
IgG2a
Clone Number
VIT200
Host
Mouse
Species Reactivity
Human
Form/Format
PBS pH 7.2, 1% BSA, 0.05% NaN3
Specificity
The CD45 mAb (clone VIT200) recognizes a pan CD45 epitope.

The sensitivity of VIT200 mAb is determined by staining well-definted blood samples from representative donors with serial-fold mAb dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI (mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50 ul of leukocytes containing 10^7 cells/ml are stained with 20 ul mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
Preparation and Storage
Monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody. For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8 degree C (DO NOT FREEZE!) and protected from prolonged exposure to light. Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.
Molecular Weight
130,898 Da
OMIM
gene 608971
CE Mark
CE
Applications Tested/Suitable for anti-CD45 antibody
Direct Immunofluorescence (IF), Indirect Immunofluorescence (IF), Flow Cytometry (FC/FACS)
Application Notes for anti-CD45 antibody
Direct Immunofluorescence (Staining Procedure) Fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations.

Proposed staining procedure for whole blood in short:
- For each sample add 50 ul of EDTA anti-coagulated blood to a 3-5 ml tube
- Add 20 ul of the appropriate monoclonal antibody conjugate
- Incubate the tube for 15 minutes at 4 degree C or at room temperature in the dark
- Add 100 ul NM-LYSE to each tube and incubate for 10 minutes at room temperature
- Add 3-4 ml of destilled water and vortex, incubate for 5-10 minutes at room temperature
- Centrifuge tube for 5 minutes at 300 g
- Aspirate supernatant and resuspend pellet in 0.3 ml of sheath fluid
- Analyze immediately or store samples at 2-8 degree C in the dark and analyze within 24 hours
Proposed staining procedure for MNC in short:
- Carefully add 20 ul antibody conjugate and 50-100 ul MNC to the bottom of a tube
- Vortex at low speed for 1-2 seconds
- Incubate for 15-30 minutes at 2-8 degree C or at room temperature
- Centrifuge tubes for 5 minutes at 300 g
- Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge cells again for 5 minutes at 300 g
- Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1 % formaldehyde and store them at 2-8 degree C in the dark. Analyze fixed cells within 24 hours

Indirect Immunofluorescence (Staining Procedure)
- Mix 20 ul purified antibody with 50 ul whole blood or MNC suspension
- Incubate for 15 minutes at 2-8 degree C
- Wash cells with phosphate buffered saline (PBS)
- Add to cell pellet 20 ul of affinity purified, fluorochrome labeled F(ab')2 anti mouse Ig antibodies
- Incubate for 15 minutes at 2-8 degree C
- Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining

Product References and Citations for anti-CD45 antibody

Battifora, H. & Trowbridge, I. S. (1983) Cancer 51, 816-21.
Brocklebank, A. M. & Sparrow, R. L. (2001) Cytometry 46, 254-61.
Cobbold, S., Hale, G. & Waldmann, H. (1987) In Leukocyte Typing III (Oxford University Press, Oxford-New York-Tokyo) p788-803.
Dalchau, R., Kirkley, J. & Fabre, J. W. (1980) Eur J Immunol 10, 737-44.
Nicholson, J. K., Hubbard, M. & Jones, B. M. (1996) Cytometry 26, 16-21.
Omary, M. B., Trowbridge, I. S. & Battifora, H. A. (1980) J Exp Med 152, 842-52
Schraven, B., Roux, M., Hutmacher, B. & Meuer, S. C. (1989) Eur J Immunol 19, 397-403.
Sugita, K., Majdic, O., Stockinger, H., Holter, W., Burger, R. & Knapp, W. (1987) Transplantation 43, 570-4.
Sun, T., Sangaline, R., Ryder, J., Gibbens, K., Rollo, C., Stewart, S. & Rajagopalan, C. (1997) Am J Clin Pathol 108, 152-7.
Thomas, M. L. (1989) Annu Rev Immunol 7, 339-69.

Other Notes

Small volumes of anti-CD45 antibody vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

Searchable Terms for CD45 purchase

MBS570368 is an antibody targeting CD45. Also known as Mouse anti CD45. Reactive in Human. Host: Mouse.

Additional Details

NCBI/Uniprot data below describe general gene information for CD45. It may not necessarily be applicable to this product.

NCBI GI #
NCBI GeneID
NCBI Accession #
UniProt Secondary Accession #
UniProt Related Accession #
NCBI Official Full Name
CD45, partial
NCBI Official Synonym Full Names
protein tyrosine phosphatase, receptor type, C
NCBI Official Symbol
PTPRC
NCBI Official Synonym Symbols
LCA; LY5; B220; CD45; L-CA; T200; CD45R; GP180
UniProt Protein Name
Receptor-type tyrosine-protein phosphatase C
UniProt Synonym Protein Names
Leukocyte common antigen; L-CA; T200; CD_antigen: CD45
UniProt Gene Name
PTPRC
UniProt Synonym Gene Names
CD45; L-CA
UniProt Entry Name
PTPRC_HUMAN
NCBI Protein Information
receptor-type tyrosine-protein phosphatase C; CD45 antigen; T200 glycoprotein; T200 leukocyte common antigen; protein tyrosine phosphatase, receptor type, c polypeptide

NCBI Summary for CD45

The protein encoded by this gene is a member of the protein tyrosine phosphatase (PTP) family. PTPs are known to be signaling molecules that regulate a variety of cellular processes including cell growth, differentiation, mitosis, and oncogenic transformation. This PTP contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains, and thus is classified as a receptor type PTP. This PTP has been shown to be an essential regulator of T- and B-cell antigen receptor signaling. It functions through either direct interaction with components of the antigen receptor complexes, or by activating various Src family kinases required for the antigen receptor signaling. This PTP also suppresses JAK kinases, and thus functions as a regulator of cytokine receptor signaling. Alternatively spliced transcripts variants of this gene, which encode distinct isoforms, have been reported. [provided by RefSeq, Jun 2012]
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UniProt Comments for CD45

CD45: a receptor type protein tyrosine phosphatase protein. Contains an extracellular domain, a single transmembrane segment and two tandem intracytoplasmic catalytic domains. The first PTPAse domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Specifically expressed in hematopoietic cells. An essential regulator of T- and B-cell antigen receptor signaling. Functions through either direct interaction with components of the antigen receptor complexes, or by activating various Src family kinases required for the antigen receptor signaling. Suppresses JAK kinases, and thus functions as a regulator of cytokine receptor signaling. Four alternatively spliced isoforms have been described.

Protein type: Membrane protein, integral; EC 3.1.3.48; Motility/polarity/chemotaxis; Receptor protein phosphatase, tyrosine

Chromosomal Location of Human Ortholog: 1q31-q32

Cellular Component: focal adhesion; membrane; integral to plasma membrane; plasma membrane; lipid raft; external side of plasma membrane

Molecular Function: protein binding; transmembrane receptor protein tyrosine phosphatase activity; protein tyrosine phosphatase activity; protein kinase binding

Biological Process: B cell proliferation; axon guidance; substrate-bound cell migration, cell release from substrate; regulation of cell cycle; protein amino acid dephosphorylation; positive regulation of antigen receptor-mediated signaling pathway; T cell receptor signaling pathway; bone marrow development; B cell receptor signaling pathway; stem cell development; cell surface receptor linked signal transduction; dephosphorylation; release of sequestered calcium ion into cytosol; positive regulation of protein kinase activity; negative regulation of T cell mediated cytotoxicity; positive regulation of B cell proliferation; negative regulation of protein kinase activity; hemopoietic progenitor cell differentiation; positive regulation of T cell proliferation; negative regulation of cytokine and chemokine mediated signaling pathway; immunoglobulin biosynthetic process; defense response to virus; T cell differentiation

Disease: Hepatitis C Virus, Susceptibility To; Severe Combined Immunodeficiency, Autosomal Recessive, T Cell-negative, B Cell-positive, Nk Cell-positive
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Precautions

All of MyBioSource's Products are for scientific laboratory research purposes and are not for diagnostic, therapeutics, prophylactic or in vivo use. Through your purchase, you expressly represent and warrant to MyBioSource that you will properly test and use any Products purchased from MyBioSource in accordance with industry standards. MyBioSource and its authorized distributors reserve the right to refuse to process any order where we reasonably believe that the intended use will fall outside of our acceptable guidelines.

Disclaimer

While every efforts were made to ensure the accuracy of the information provided in this datasheet, MyBioSource will not be liable for any omissions or errors contained herein. MyBioSource reserves the right to make changes to this datasheet at any time without prior notice.

It is the responsibility of the customer to report product performance issues to MyBioSource within 30 days of receipt of the product. Please visit our Terms & Conditions page for more information.

Product Note

Select online data sheet information is drawn from bioinformatics databases, occasionally resulting in ambiguous or non-relevant product information. It is the responsibility of the customer to review, verify, and evaluate the information to make sure it matches their requirements before purchasing the kit. Our ELISA Kit assays are dynamic research tools and sometimes they may be updated and improved. If the format of this assay is important to you then please request the current manual or contact our technical support team with a presales inquiry before placing an order. We will confirm the current details of the assay. We cannot guarantee the sample manual posted online is the most current manual, it is intended to serve as an example only. Please refer to the instructions For Use provided with the assay kit for precise details.

Product PubMed Publications

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