Description
This 5C3 monoclonal antibody specifically binds to CD40, a 45-48 kDa type I integral membrane glycoprotein. CD40 is expressed on B lymphocytes, but is not expressed on terminally differentiated B cells. CD40 is also expressed by endothelial cells, basal epithelial cells and some epithelial cell carcinomas, follicular dendritic cells, macrophages, fibroblasts, keratinocytes, and CD34+ hematopoietic progenitor cells. This antibody is useful for studying the roles played by CD40 in B-cell growth, proliferation, and differentiation including immunoglobulin isotype switching. Anti-CD40 antibodies have been reported to stimulate B-cell proliferation when costimulated with anti-µ, anti-CD20 antibodies or with phorbol esters. 5C3 is capable of inducing B-cell proliferation when presented with IL-4.
Clone 5C3 reacts with the human form of the 45-48 kDa type I integral membrane glycoprotein, CD40. This clone also cross-reacts with a subset of peripheral blood lymphocytes, but not monocytes nor granulocytes, of baboon and both rhesus and cynomolgus macaque monkeys. The distribution on lymphocytes is similar to that seen with normal human donor lymphocytes, with the reactivity being restricted to CD20+ lymphocytes.
Format
- Format
Alexa Fluor® 700
- Excitation Source
Red 633 nm
- Excitation Max
696 nm
- Emission Max
719 nm
Alexa Fluor® 700 is a far-red dye that can be excited with a 633–640-nm laser. This enables multicolor analysis in conjunction with APC or Alexa Fluor® 647, and APC-H7 or APC-Cy7 reagents.
Suggested Companion Products
Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated to Alexa Fluor® 700 under optimum conditions, and unreacted Alexa Fluor® 700 was removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Alexa Fluor® 700 has an adsorption maximum of ~700nm and a peak fluorescence emission of ~720nm. Before staining cells with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species testing during development may have been performed with a different format of the same clone. Selected applications have been tested for cross-reactivity.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
