CD45 Antibody (135-4C5)

Novus Biologicals | Catalog # NBP2-44863

Clone 135-4C5 was used by HLDA to establish CD designation.
Novus Biologicals

Key Product Details

Species Reactivity

Human

Applications

Immunohistochemistry, Immunohistochemistry-Paraffin, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2b Kappa Clone # 135-4C5
View all CD45 Primary Antibodies »

Product Specifications

Immunogen

Stimulated human leukocytes

Localization

Cell surface and cytoplasmic

Marker

Leucocyte Marker

Specificity

CD45R, also designated CD45 and PTPRC, has been identified as a transmembrane glycoprotein, broadly expressed among hematopoietic cells. Multiple isoforms of CD45R are distributed throughout the immune system according to cell type. These isoforms arise because of alternative splicing of exons 4, 5, and 6. The corresponding protein domains are characterized by the binding of monoclonal antibodies specific for CD45RA (exon 4), CD45RB (exon 5), CD45RC (exon 6) and CD45RO (exons 4 to 6 spliced out). The variation in these isoforms is localized to the extracellular domain of CD45R, while the intracellular domain is conserved. CD45R functions as a phosphor-tyrosine phosphatase. This monoclonal antibody reacts with all isoforms of CD45R expressed by all hematopoietic cells, except erythrocytes, having a higher level of expression on lymphocytes than on granulocytes (Workshop IV). Antibody to CD45 is useful in differential diagnosis of lymphoid tumors from non-hematopoietic undifferentiated neoplasms.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2b Kappa

Theoretical MW

147 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Description

200ug/ml of antibody purified from Bioreactor Concentrate by Protein A or G. Prepared in 10 mM PBS with 0.05% BSA & 0.05% azide. Also available WITHOUT BSA & azide at 1.0 mg/ml. (NBP3-11434)

Antibody with azide - store at 2 to 8C. Antibody without azide - store at -20 to -80C.

Scientific Data Images for CD45 Antibody (135-4C5)

Immunocytochemistry/ Immunofluorescence: CD45 Antibody (135-4C5) [NBP2-44863]

Immunocytochemistry/ Immunofluorescence: CD45 Antibody (135-4C5) [NBP2-44863]

Immunocytochemistry/Immunofluorescence: CD45 Antibody (135-4C5) [NBP2-44863] - Immunofluorescence Analysis of PFA-fixed Jurkat cells labeling CD45RA with CD45 antibody (135-4C5) [NBP2-44863] followed by Goat anti-Mouse IgG-CF488 (Green). The nuclear counterstain is Red Dot.
Immunohistochemistry-Paraffin: CD45 Antibody (135-4C5) [NBP2-44863]

Immunohistochemistry-Paraffin: CD45 Antibody (135-4C5) [NBP2-44863]

Immunohistochemistry-Paraffin: CD45 Antibody (135-4C5) [NBP2-44863] - Human Tonsil stained with CD45 antibody (135-4C5) [NBP2-44863].
Flow Cytometry: CD45 Antibody (135-4C5) [NBP2-44863]

Flow Cytometry: CD45 Antibody (135-4C5) [NBP2-44863]

Flow Cytometry: CD45 Antibody (135-4C5) [NBP2-44863] - Flow cytometry analysis of lymphocyte gated PBMCs unstained (gray) or stained with CD45 antibody (135-4C5) (green).
Flow Cytometry: CD45 Antibody (135-4C5) [NBP2-44863]

Flow Cytometry: CD45 Antibody (135-4C5) [NBP2-44863]

Flow Cytometry: CD45 Antibody (135-4C5) [NBP2-44863] - Flow Cytometric Analysis of PFA-fixed Jurkat cells using CD45 antibody (135-4C5) [NBP2-44863] followed by Goat anti-Mouse-IgG-CF488 (Blue); Isotype Control (Red).

Applications for CD45 Antibody (135-4C5)

Application
Recommended Usage

Flow Cytometry

1-2 ug/million cells

Immunocytochemistry/ Immunofluorescence

1-2 ug/ml

Immunohistochemistry-Paraffin

1-2 ug/ml
Application Notes
Immunohistochemistry (Formalin-fixed): 1-2ug/ml for 30 minutes at RT. Staining of formalin-fixed tissues requires heating tissue sections in 10mM Tris with 1mM EDTA, pH 9.0, for 45 min at 95C followed by cooling at RT for 20 minutes.
Optimal dilution for a specific application should be determined.

Flow Cytometry Panel Builder

Bio-Techne Knows Flow Cytometry

Save time and reduce costly mistakes by quickly finding compatible reagents using the Panel Builder Tool.

Advanced Features

  • Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
  • Spillover Popups - Visualize the spectra of individual fluorochromes
  • Antigen Density Selector - Match fluorochrome brightness with antigen density
Build Your Panel Now
Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

10 mM PBS with 0.05% BSA

Preservative

0.05% Sodium Azide

Concentration

0.2 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C.

Background: CD45

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

Long Name

Cluster of Differentiation 45

Alternate Names

CD45, LCA, PTPRC, T200 Glycoprotein

Gene Symbol

PTPRC

UniProt

P08575

Additional CD45 Products

Product Documents for CD45 Antibody (135-4C5)

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Download SDS

Product Specific Notices for CD45 Antibody (135-4C5)

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.

Citations for CD45 Antibody (135-4C5)

Customer Reviews for CD45 Antibody (135-4C5)

There are currently no reviews for this product. Be the first to review CD45 Antibody (135-4C5) and earn rewards!

Have you used CD45 Antibody (135-4C5)?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

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

FAQs for CD45 Antibody (135-4C5)

Showing  1 - 5 of 5 FAQs Showing All

    • Q: Could you please let me know the supply form (Liquid or Lyohpilized) of this antibody? 
      A: This antibody is provided in 10 mM PBS with 0.05% BSA & 0.05% azide; it is in the liquid format. The concentration is 200 ug/ml, regardless the package sizes (0.02 mg, 0.1, and 0.2 mg).

    • Q: For use in Western Blot with CD45 antibodies, what molecular weight of the band should I expect to see?
      A: 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.

    • Q: 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?
      A: 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.

    • A: 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.

    • Q: 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?
      A: 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.
Showing  1 - 5 of 5 FAQs Showing All
View all FAQs for Antibodies