Alexa Fluor® 647 Mouse Anti-Pig CD4a
Clone 74-12-4 (RUO)
- Brand BD Pharmingen™
- Alternative Name CD4; CD4 molecule; Lymphocyte antigen CD4
- Concentration 0.2 mg/ml
- Isotype Mouse BALB/c IgG2b, κ
- Reactivity Pig (QC Testing)
Flow cytometry (Routinely Tested)
- Immunogen dd miniature swine thymocytes
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The 74-12-4 (also known as clone PT4) monoclonal antibody specifically binds to CD4, a 55-kDa antigen expressed on T lymphocytes. This antibody does not react with CTL effectors, CTL precursors, or NK cells (ie, CD8[bright] cells) and it does not cross-react with human or bovine cells. Two peripheral T-helper lymphocyte phenotypes can be distinguished in the pig: CD4+CD8- and CD4+CD8[dull]. mAb 74-12-4 has been reported to inhibit proliferative responses of peripheral blood lymphocytes to mitogen, soluble antigen, and alloantigen. It is only marginally effective for in vivo depletion of peripheral CD4+ T cells. Two alloantigenic forms of CD4 have been recognized in miniature swine based upon their recognition (CD4.1) or lack of recognition (CD4.2) by mAb 74-12-4; the CD4.2 phenotype displays an autosomal recessive, non-MHC-linked, pattern of inheritance. The molecular basis for the polymorphism is a cluster of nucleotide differences leading to multiple amino-acid substitutions in the Ig CDR2-like loop structure. This mAb was clustered as anti-CD4a at the First International Swine CD Workshop. It has been reported to crossreact with chicken leukocytes.
Alexa Fluor® 647 conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC. However, APC tends to be brighter while Alexa Fluor® 647 is more optimal for intracellular applications. This fluorochrome exhibits uncommon photostability, making it an ideal choice for use in fluorescence microscopy. Due to nearly identical excitation and emission properties but different spillover characteristics, APC and Alexa Fluor® 647 cannot be used simultaneously.
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Preparation and Storage
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to www.bdbiosciences.com/pharmingen/protocols for technical protocols.
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.