BB515 Mouse Anti-Human CD3
Clone HIT3a (RUO)
- Brand BD Horizon™
- Alternative Name CD3E; T3E; TCRE; cd 3; cd-3; cd3; CD3-epsilon; 916
- Vol. Per Test 5 µl
- Isotype Mouse IgG2a, κ
- Reactivity Human (QC Testing)
Flow cytometry (Routinely Tested)
- Workshop No. V 5T-CD03.05
- Storage Buffer Aqueous buffered solution containing ≤0.09% sodium azide.
- Regulatory Status RUO
Regulatory Status Legend
The HIT3a monoclonal antibody specifically binds to the human CD3ε-chain, a 20 kDa subunit of the CD3/T cell antigen receptor complex found on 70-80% of normal human peripheral blood lymphocytes and 60-85% of thymocytes. Studies from the HLDA Workshop show that this antibody can be mitogenic for T lymphocytes. The CD3 complex plays a role in signal transduction during antigen recognition by the T cell receptor. HIT3a antibody does not stain intracellular CD3 unlike the other CD3-specific clone, UCHT1 (Cat. No. 555330/550368).
The antibody was conjugated to BD Horizon BB515 which was developed exclusively by BD Biosciences. With an excitation max of 490 nm and an emission max of 515 nm, BD Horizon BB515 can be excited by the 488 nm laser and detected in a standard FITC set (eg, 530/30-nm filter). This dye provides a much brighter alternative to FITC with less spillover into the PE detector.
BB515 is a dye that was exclusively developed by BD Biosciences as a brighter alternative to FITC. This dye is up to seven times brighter than FITC and has less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously.
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Preparation and Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
The antibody was conjugated with BD Horizon™ BB515 under optimum conditions and unconjugated antibody was removed.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.