Analysis Note

ControlJurkat cell lysate

Application

Research Sub CategoryImmunological Signaling

Research CategorySignaling

Anti-CD45 Antibody, clone F10-89-4 detects level of CD45 & has been published & validated for use in FC, WB, IF, IP, IH(P).

Immunofluorescence: A previous lot of this antibody was used in the identification of cells of leucocyte origin by indirect staining.

Immunohistochemistry (paraffin): A previous lot of this antibody was used in the discrimination between malignant cells of haematopoietic origin and other malignancies in frozen tissue sections.

Immunoprecipitation: A previous lot of this antibody was used in studies of the leucocyte common antigen complex and its identification by the Western blotting technique.

Flow Cytometry: Studies of CD45 expression and the association with T-cell receptor signaling was performed using this antibody of previous lot.
Studies of the mechanism by which CD45 regulates B cell activation through protein tyrosine phospatases.
Optimal working dilutions must be determined by the end user.

Immunohistochemistry(paraffin): Representative testing from a previous lot.

Optimal Staining of CD45 Monoclonal Antibody: Tonsil

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

CD45 is a receptor-tyrosine phosphatase required for antigen-induced signaling in T-cells and B-cells; its substrates are unknown. Interestingly, it associates with several Src-family tyrosine kinases, which are inhibited by tyrosine phosphorylation in their c-termini, suggesting a mechanism for signaling by CD45.

Immunogen

Purified T cells from human lymph nodes

Linkage

Replaces: 04-1102

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified mouse monoclonal IgG2a presented in phosphate buffered saline containing 10 mM sodium azide and 1 mg/mL bovine serum albumin.
We recommend that each laboratory determine an optimum working titre for use in its particular application.

Protein A affinity chromatography

Quality

Routinely evaluated by Western Blot on jurkat cell lysate.

Western Blot Analysis: 1:500 dilution of this lot detected CD45 on 10 µg of Jurkat lysates

Specificity

This antibody reacts with the leucocyte common antigen (LCA) complex to be found on lymphocytes, monocytes, granulocytes, thymocytes and malignant T and B cells. No reactivity has been observed with primary or metastatic carcinoma cells. Plasma cells or myeloma cells may have weak expression or be negative for this antigen.
The LCA comprises of at least 5 isoforms ranging from 180-220 kDa which are produced as a result of alternative splicing of exons A, B or C. CBL 124 recognizes epitopes common to
all of the isoforms.
Antigen distribution:
Thymocytes >95%
Granulocytes >95%
Monocytes >95%
B cells (CD20+) >95%
T cells (CD3+) >95%
NK cells (CD16+) >95%
Peripheral blood lymphocytes >95%
FUSION PARTNER: NS1 myeloma cell line

Storage and Stability

For use within 1 month of purchase store at 2-8°C, for long term storage aliquot antibody into small volumes and store at -20°C for up to one year from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Target description

~180-220 kDa