CD45 Antibody (MEM-28) [DyLight 680]

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Flow Cytometry: CD45 Antibody (MEM-28) [DyLight 680] [NB500-319FR] - Surface staining of human peripheral blood cells with anti-human CD45 antibody (MEM-28) FITC.

Product Details

Summary
Reactivity Hu, Mu, Eq(-)Species Glossary
Applications WB, Flow, ICC/IF, IHC, IHC-P
Clone
MEM-28
Clonality
Monoclonal
Host
Mouse
Conjugate
DyLight 680

CD45 Antibody (MEM-28) [DyLight 680] Summary

Immunogen
This CD45 Antibody (MEM-28) was prepared from Human thymocytes and T lymphocytes.
Localization
Type I membrane protein
Specificity
CD45 Antibody (MEM-28) reacts with all alternative forms of human CD45 antigen (Leukocyte Common Antigen), a 180-220 kDa single chain type I transmembrane protein expressed at high level on all cells of hematopoietic origin, except erythrocytes and platelets. HLDA III; WS Code NL 833a
Isotype
IgG1
Clonality
Monoclonal
Host
Mouse
Gene
PTPRC
Purity
Protein A purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot
  • Flow Cytometry
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry
  • Immunohistochemistry-Paraffin
Application Notes
Optimal dilution of this antibody should be experimentally determined.

Reactivity Notes

Mouse reactivity reported in scientific literature (PMID: 27272386). Please note that CD45 Antibody (MEM-28) is reactive to Mouse and derived from the same host, Mouse. Mouse-On-Mouse blocking reagent may be needed for IHC and ICC experiments to reduce high background signal. You can find these reagents under catalog numbers PK-2200-NB and MP-2400-NB. Please contact Technical Support if you have any questions.

Packaging, Storage & Formulations

Storage
Store at 4C in the dark.
Buffer
50mM Sodium Borate
Preservative
0.05% Sodium Azide
Purity
Protein A purified

Notes



DyLight (R) is a trademark of Thermo Fisher Scientific Inc. and its subsidiaries.

Alternate Names for CD45 Antibody (MEM-28) [DyLight 680]

  • 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, also known as leukocyte common antigen (LCA), T200, or Ly5, is a member C of the class 1 receptor-like protein tyrosine phosphatase (PTPRC) family (1, 2). It is a transmembrane glycoprotein which, due to alternative splicing, has a multiple isoforms with a theoretical molecular weight ranging from 180 - 220 kDa (1, 3-5). Human CD45 is synthesized as a 1281 amino acid sequence consisting of an alternatively spliced extracellular receptor-like region, a cysteine-rich domain, fibronectin-like III repeats, a transmembrane segment, and a cytoplasmic region with tandem protein tyrosine phosphatase (PTP) domains: the membrane proximal domain (D1) and the membrane distal domain (D2) (3, 5). CD45 is expressed on all nucleated hematopoietic cells and their precursors, except mature red blood cells, and is one of the most abundantly-expressed cell-surface glycoproteins, comprising approximately 10% of surface proteins in lymphocytes (3). Functionally, CD45 is essential for development and activation of T-cells and B-cells (1-5). More specifically, CD45 positively regulates antigen receptor signaling and Src-family member kinase activity (1, 3). There are many ways to regulate CD45 phosphatase activity including ligand binding, dimerization, protein interactions, cellular localization, and covalent modifications (3, 6). Ligands for CD45 include pUL11, a transmembrane protein of the cytomegalovirus RL11 (CMV RL11) family, and placental protein 14 (PP14), both of which exclusively bind CD45, and various lectins including CD22, galectin-1, galectin-3, macrophage mannose receptor (MR), and macrophage galactose-type lectin (MGL) (6).

Given its role in immune cell development and activation, CD45 has also been linked to a variety of diseases. The importance of CD45 in immunity has been revealed in human and mouse studies where CD45-deficiency leads to a severe-combined immunodeficiency (SCID) phenotype (2, 3, 6). A CD45-knockout mice study revealed inhibited thymocyte production and poor B-cell response, whereas CD45 activation in mice causes lymphoproliferation and autoantibody production (3). CD45 variants have been associated with altered immune function and autoimmune disorders including multiple sclerosis, systemic lupus erythematosus (SLE), and rheumatoid arthritis (6). Furthermore, altered CD45 expression has been implicated in oncological conditions including chronic lymphatic leukemia, acute lymphatic leukemia, Hodgkin lymphoma, multiple myeloma, and diffuse large B-cell lymphoma (6). Considering its role in autoimmune disorders, immunodeficiency and cancer, CD45 is an ideal therapeutic target (3, 6). The main approaches to control CD45 function is through either selective inhibitors or anti-CD45 antibodies (3).

Alternative names for CD45 includes B220, CD antigen: CD45, CD45 antigen, CD45R, EC 3.1.3.48, GP180, LCA, Leukocyte common antigen, LY5, protein tyrosine phosphatase receptor type c polypeptide, PTPRC, receptor-type tyrosine-protein phosphatase C, T200 Glycoprotein, and T200.

References

1. Trowbridge, I. S., & Thomas, M. L. (1994). CD45: an emerging role as a protein tyrosine phosphatase required for lymphocyte activation and development. Annual review of immunology. https://doi.org/10.1146/annurev.iy.12.040194.000505

2. Andersen, J. N., Jansen, P. G., Echwald, S. M., Mortensen, O. H., Fukada, T., Del Vecchio, R., Tonks, N. K., & Moller, N. P. (2004). A genomic perspective on protein tyrosine phosphatases: gene structure, pseudogenes, and genetic disease linkage. FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

3. Hermiston, M. L., Xu, Z., & Weiss, A. (2003). CD45: a critical regulator of signaling thresholds in immune cells. Annual review of immunology. https://doi.org/10.1146/annurev.immunol.21.120601.140946

4. Tonks, N. K., Diltz, C. D., & Fischer, E. H. (1990). CD45, an integral membrane protein tyrosine phosphatase. Characterization of enzyme activity. The Journal of biological chemistry.

5. Nam, H. J., Poy, F., Saito, H., & Frederick, C. A. (2005). Structural basis for the function and regulation of the receptor protein tyrosine phosphatase CD45. The Journal of experimental medicine. https://doi.org/10.1084/jem.20041890

6. Rheinlander, A., Schraven, B., & Bommhardt, U. (2018). CD45 in human physiology and clinical medicine. Immunology letters. https://doi.org/10.1016/j.imlet.2018.01.009

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.

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Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for CD45 Antibody (NB500-319FR). (Showing 1 - 4 of 4 FAQ).

  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.

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB500-319AF350
Alexa Fluor 405 NB500-319AF405
Alexa Fluor 488 NB500-319AF488
Alexa Fluor 532 NB500-319AF532
Alexa Fluor 594 NB500-319AF594
Alexa Fluor 647 NB500-319AF647
Alexa Fluor 700 NB500-319AF700
Alexa Fluor 750 NB500-319AF750
Allophycocyanin NB500-633
Biotin NB110-81718
DyLight 350 NB500-319UV
DyLight 405 NB500-319V
DyLight 488 NB500-319G
DyLight 550 NB500-319R
DyLight 594 NB500-319DL594
DyLight 650 NB500-319C
DyLight 680 NB500-319FR
DyLight 755 NB500-319IR
FITC NB110-81719
HRP NB500-319H
Janelia Fluor 549 NB500-319JF549
Janelia Fluor 646 NB500-319JF646
PE NBP1-44756
PerCP NB110-81720

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Bioinformatics Tool for CD45 Antibody (NB500-319FR)

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By Jennifer Sokolowski, MD, PhD.Gut-microbiome interactionsOn and in our bodies, microbes outnumber our cells by about ten-to-one. Studies have revealed that the microbiome influences neurogenesis, cognition, and...  Read full blog post.

How to identify B cell subsets using flow cytometry
By Victoria OsinskiUsing flow cytometry to identify B cell subsetsIdentifying cellular subsets by flow cytometry requires careful and thorough planning in order to ensure the correct subset of cells are identified a...  Read full blog post.

You complete me: Natural killer cells need TGF-beta inhibition to effectively combat cancers
By Jamshed Arslan Pharm.D. Natural killer (NK) cells are lymphocytes of the innate immune system that were first discovered for their “natural” ability to kill cancer cells. To use NK cells as anti-cancer therapy, t...  Read full blog post.

The application of CD31/Pecam-1 (MEC 7.46) in breast cancer research
CD31/PECAM-1, or platelet endothelial cell adhesion molecule 1, is a 130-kDa glycoprotein expressed on vascular and hematopoietic cells.  Depending on the cell type, CD31/PECAM-1 expression can be largely localized to cell junctions, playing a rol...  Read full blog post.

CD45 - Much more than just a housekeeping protein
CD45, also known as T200 or the Leukocyte Common Antigen (LCA), is encoded by the Protein Tyrosine Phosphatase Receptor type C (PTPRC) gene. The protein is expressed exclusively on cells of the haematopoietic system, and is one of the most abundant le...  Read full blog post.

CD45 Isoforms: Hematopoietic Differentiation, Cancer and Alzheimer's
CD45, also known as protein tyrosine phosphatase, receptor type, C (PTPRC), was originally known as common leukocyte antigen and is a signal transducer involved in many physiological processes such as growth and differentiation, cancer transformation,...  Read full blog post.

Vimentin Antibodies in Rheumatoid Arthritis & Cataracts Research
Vimentin is a 57kDa type III intermediate filament (IF) protein that is the major cytoskeletal component of mesenchymal cells and the first to be expressed during cell differentiation. It plays a significant role in supporting and anchoring the positi...  Read full blog post.

Antibodies In The Differential Diagnosis Of Undifferentiated Tumors
Immunohistochemical testing using antibody panels has a valuable role in cancer diagnosis. Some tumors, especially more malignant ones, tend to lose the appearance of the tissue of origin. However, knowing the tissue of origin assists the physician in...  Read full blog post.

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Bioinformatics

Gene Symbol PTPRC