Human CD45 Antibody

Catalog #: MAB14301 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB14301-100
MAB14301-SP

Detection of CD45 in Human Blood Lymphocytes by Flow Cytometry.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human CD45 Antibody Summary

Species Reactivity

Human

Specificity

Detects human CD45 in direct ELISAs.

Source

Monoclonal Mouse IgG₁ Clone # 998209

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant human CD45
Thr24-Asn575
Accession # P08575

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Flow Cytometry

0.25 µg/10⁶ cells

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

5-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Flow Cytometry

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Detection of CD45 in Human Blood Lymphocytes by Flow Cytometry. Human peripheral blood lymphocytes were stained with Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB14301, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). View our protocol for Staining Membrane-associated Proteins.

Immunocytochemistry

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CD45 in Human PBMCs and MCF-7 Cell Line. CD45 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs; positive staining, left panel) and MCF-7 human breast cancer cell line (negative staining, right panel) using Mouse Anti-Human CD45 Monoclonal Antibody (Catalog # MAB14301) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: CD45

CD45, previously called LCA (leukocyte common antigen), T200, or Ly5 in mice, is member C of the class 1 (receptor‑like) protein tyrosine phosphatase family (PTPRC) (1, 2). It is a variably glycosylated 180‑220 kDa transmembrane protein that is abundantly expressed on all nucleated cells of hematopoietic origin (1‑3). CD45 has several isoforms, expressed according to cell type, developmental stage and antigenic exposure (1‑5). The longest form, CD45RABC (called B220 in mouse), is expressed on B lymphocytes (5). The CD45RABC cDNA encodes 1304 amino acids (aa), including a 23 aa signal sequence, a 552 aa extracellular domain containing the splicing region, a cysteine‑rich region and two fibronectin type III domains, a 22 aa transmembrane sequence, and a 707 aa cytoplasmic domain that contains two phosphatase domains, D1 and D2. Only D1 has phosphatase activity. CD45R0 is the shortest form, lacking exons 4, 5 and 6 which encode aa 32‑191. It is expressed on memory cells, while intermediate sizes are expressed on other T cells (3, 4, 6). CD45 has been best studied in T cells, where it determines T cell receptor signaling thresholds (3, 6‑8). CD45 is moved into or out of the immunological synapse (IS) membrane microdomain depending on the relative influence of interaction with the extracellular galectin lattice or the intracellular actin cytoskeleton (9, 10). Galectin interaction can be fine‑tuned by varying usage of the heavily O‑glycosylated spliced regions and sialylation of N‑linked carbohydrates (4, 9). Within the IS, CD45 dephosphorylates and negatively regulates the Src family kinase, Lck (8‑10). In other leukocytes, CD45 influences differentiation and links immunoreceptor signaling with cytokine secretion and cell survival, partially overlapping in function with DEP‑1/CD148 (11‑14). CD45 deletion causes in severe immunodeficiency, while point mutations may be associated with autoimmune disorders (6, 7).

References

Anderson, J.N. et al. (2004) FASEB J. 18:8.
Streuli, M. et al. (1987) J. Exp. Med. 166:1548.
Hermiston, M.L. et al. (2003) Annu. Rev. Immunol. 21:107.
Earl, L.A. and L.G. Baum (2008) Immunol. Cell Biol. 86:608.
Ralph, S.J. et al. (1987) EMBO J. 6:1251.
Falahti, R. and D. Leitenberg (2008) J. Immunol. 181:6082.
Tchilian, E.Z. and P.C.L. Beverley (2006) Trends Immunol. 27:146.
McNiell, L. et al. (2007) Immunity 27:425.
Chen, I-J. et al. (2007) J. Biol. Chem. 282:35361.
Freiberg, B.A. et al. (2002) Nat. Immunol. 3:911.
Zhu, J.W. et al. (2008) Immunity 28:183.
Huntington, N.D. et al. (2006) Nat. Immunol. 7:190.
Hesslein, D.G. et al. (2006) Proc. Natl. Acad. Sci. USA 103:7012.
Cross, J.L. et al. (2008) J. Immunol. 180:8020.

Long Name

Cluster of Differentiation 45

Entrez Gene IDs

5788 (Human); 19264 (Mouse); 490255 (Canine); 100061950 (Equine)

Alternate Names

B220; CD_antigen: CD45; CD45 antigen; CD45; CD45R; EC 3.1.3.48; EC:3.1.3.48; GP180; LCA; L-CA; Leukocyte common antigen; LY5; protein tyrosine phosphatase, receptor type, c polypeptide; PTPRC; receptor-type tyrosine-protein phosphatase C; T200 Glycoprotein; T200 leukocyte common antigen; T200