CD45 Antibody (2D1) [Biotin]

CD45 was detected in immersion fixed human peripheral blood mononuclear cells (PBMCs) using Mouse Anti-Human CD45 Biotinylated Monoclonal Antibody (Catalog # BAM1430) at 25 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Streptavidin (red; Catalog # NL999) and counterstained with DAPI (blue). Specific staining was localized to plasma membrane. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

• Reactivity: Hu
• Applications: Flow, ICC/IF
• Clone: 2D1
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Biotin
• Concentration: LYOPH

CD45 Antibody (2D1) [Biotin] Summary

• Immunogen: Human peripheral blood mononuclear cells
• Specificity: Detects human CD45. This antibody recognizes all isoforms of human CD45.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: PTPRC
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunocytochemistry 8-25 ug/mL
• Readout System:
  • Streptavidin Native Protein NBP2-29367
  • Streptavidin Native Protein NBP2-29370
  • Streptavidin Native Protein NBP2-29371

Packaging, Storage & Formulations

• 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.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for CD45 Antibody (2D1) [Biotin]

• 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

Background

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).

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

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for CD45 Antibody (BAM1430)(12)

Publications using BAM1430

• Hongshan Guo, Joanna A. Vuille, Ben S. Wittner, Emily M. Lachtara, Yu Hou, Maoxuan Lin et al. DNA hypomethylation silences anti-tumor immune genes in early prostate cancer and CTCs Cell 2023-06-22 [PMID: 37327786]
• RY Ebright, MA Zachariah, DS Micalizzi, BS Wittner, KL Niederhoff, LT Nieman, B Chirn, DF Wiley, B Wesley, B Shaw, E Nieblas-Be, L Atlas, A Szabolcs, AJ Iafrate, M Toner, DT Ting, PK Brastianos, DA Haber, S Maheswaran HIF1A signaling selectively supports proliferation of breast cancer in the brain Nature Communications, 2020-12-09;11(1):6311. 2020-12-09 [PMID: 33298946] (Neutralization, Human)
• JW Franses, J Philipp, P Missios, I Bhan, A Liu, C Yashaswini, E Tai, H Zhu, M Ligorio, B Nicholson, EM Tassoni, N Desai, AS Kulkarni, A Szabolcs, TS Hong, AS Liss, C Fernandez-, DP Ryan, S Maheswaran, DA Haber, GQ Daley, DT Ting Pancreatic circulating tumor cell profiling identifies LIN28B as a metastasis driver and drug target Nat Commun, 2020-07-03;11(1):3303. 2020-07-03 [PMID: 32620742] (Neutralization, Human)
• Hakan Inan, James L. Kingsley, Mehmet O. Ozen, Huseyin Cumhur Tekin, Christian R. Hoerner, Yoriko Imae et al. Monitoring Neutropenia for Cancer Patients at the Point of Care Small Methods 2017-09-11 [PMID: 30740513] (Cell Capture, Human)
• Nicola Aceto, Aditya Bardia, Ben S. Wittner, Maria C. Donaldson, Ryan O'Keefe, Amanda Engstrom et al. AR Expression in Breast Cancer CTCs Associates with Bone Metastases Molecular Cancer Research 2018-04-01 [PMID: 29453314]
• X Hong, RJ Sullivan, M Kalinich, TT Kwan, A Giobbie-Hu, S Pan, JA LiCausi, JD Milner, LT Nieman, BS Wittner, U Ho, T Chen, R Kapur, DP Lawrence, KT Flaherty, LV Sequist, S Ramaswamy, DT Miyamoto, M Lawrence, M Toner, KJ Isselbache, S Maheswaran, DA Haber Molecular signatures of circulating melanoma cells for monitoring early response to immune checkpoint therapy Proc. Natl. Acad. Sci. U.S.A., 2018-02-16;0(0):. 2018-02-16 [PMID: 29453278] (Cell Selection, Human)
• Joseph W. Franses, Omer Basar, Abdurrahman Kadayifci, Osman Yuksel, Melissa Choz, Anupriya S. Kulkarni et al. Improved Detection of Circulating Epithelial Cells in Patients with Intraductal Papillary Mucinous Neoplasms The Oncologist 2018-01-01 [PMID: 28860411] (Bioassay, Human)
• Y Zheng, DT Miyamoto, BS Wittner, JP Sullivan, N Aceto, NV Jordan, M Yu, NM Karabacak, V Comaills, R Morris, R Desai, N Desai, E Emmons, JD Milner, RJ Lee, CL Wu, LV Sequist, W Haas, DT Ting, M Toner, S Ramaswamy, S Maheswaran, DA Haber Expression of ?-globin by cancer cells promotes cell survival during blood-borne dissemination Nat Commun, 2017-02-09;8(0):14344. 2017-02-09 [PMID: 28181495] (Bioassay, Human)
• HER2 expression identifies dynamic functional states within circulating breast cancer cells Nature, 2016-08-24;0(0):. 2016-08-24 [PMID: 27556950] (Flow Cytometry, Human)
• Tseng C, Biancotti J, Berg B, Gate D, Kolar S, Muller S, Rodriguez M, Rezai-Zadeh K, Fan X, Beenhouwer D, Town T, Liu G Increased Susceptibility of Humanized NSG Mice to Panton-Valentine Leukocidin and Staphylococcus aureus Skin Infection. PLoS Pathog, 2015-11-30;11(11):e1005292. 2015-11-30 [PMID: 26618545] (IHC-P, Human)
• David T. Ting, Ben S. Wittner, Matteo Ligorio, Nicole Vincent Vincent Jordan, Ajay M. Shah, David T. Miyamoto et al. Single-Cell RNA Sequencing Identifies Extracellular Matrix Gene Expression by Pancreatic Circulating Tumor Cells Cell Reports 2014-09-25 [PMID: 25242334]
• Brownell J, Wagoner J, Lovelace E, Thirstrup D, Mohar I, Smith W, Giugliano S, Li K, Crispe I, Rosen H, Polyak S Independent, parallel pathways to CXCL10 induction in HCV-infected hepatocytes. J Hepatol, 2013-06-12;59(4):701-8. 2013-06-12 [PMID: 23770038] (Cell Selection, Human)

FAQs for CD45 Antibody (BAM1430). (Showing 1 - 4 of 4 FAQs).

1. We would like to order a CD45 antibody to stain immune cells that were isolated from a ligated sciatic nerve of mice with double immunofluorescence (using PFA-fixed tissue on slides). Which of the following rat monoclonal CD45 antibodies would you recommend: 30-F11, IBL-3/16 or 5C16?
   • I would recommend clone 30-F11. As clone IBL-3/16 has not yet been validated for IHC-P application, we would not be able to guarantee its workability on PFA-fixed tissue. Although both other clones i.e. 30-F11 and 5C16, would be good for your samples, CD45 30-F11 # NB100-77417 is a well known clone that offers more flexibility around protocol because of availability of its conjugated forms. It would be advantageous to use a conjugated primary as you are planning for double-immunostaining procedure.
2. For use in Western Blot with CD45 antibodies, what molecular weight of the band should I expect to see?
   • The theoretical molecular weight for most of our CD45 antibodies is 147 kDa based off the first isoform. Any variation on 147 is due to the immunogen being from a different species and the protein being a slightly different size. CD45 is a family of single chain transmembraneous glycoproteins consisting of at least four isoforms (220, 205, 190, 180 kDa) which share a common large intracellular domain. Their extracellular domains are heavily glycosylated.
3. If this product is used in an application or species as a part of a customer review, will that validate this product in the application/species?
   • If any of our primary antibodes are used in an untested application or species and it is shown to work through images from customer reviews or through publications, this validates the application/species for this product, allowing the tested application/species to fall under our 100% guarantee. Please check out our Innovator's Reward Program if you decide to test a primary antibody with a species or application that is not currently listed. Please note that the Innovator's Reward Program only applies to our primary antibodies.
4. Is CD45 a good target for use in Neurodegeneration studies?
   • Yes, here are the approved research areas we have listed for our CD45 products: Adaptive Immunity, Cell Biology, Cellular Markers, Cytokine Research, Glia Markers, Hematopoietic Stem Cell Markers, Immunology, Innate Immunity, Mast Cell Markers, Mesenchymal Stem Cell Markers, Microglia Markers, Myeloid Cell Markers, Myeloid-derived Suppressor, Neurodegeneration, Neuroscience, Signal Transduction, Stem Cell Markers, Growth and Development.

Bioinformatics

• Gene Symbol: PTPRC