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Biotin Mouse Anti-Human CD3
Biotin Mouse Anti-Human CD3
Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with either Biotin Mouse IgG2a, κ Isotype Control (Cat. No. 555572; dashed line histogram) or Biotin Mouse Anti-Human CD3 antibody (Cat. No. 555338; solid line histogram), followed by Streptavidin-PE (Cat. No. 554061). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Flow cytometric analysis of CD3 expression on human peripheral blood lymphocytes. Human whole blood was stained with either Biotin Mouse IgG2a, κ Isotype Control (Cat. No. 555572; dashed line histogram) or Biotin Mouse Anti-Human CD3 antibody (Cat. No. 555338; solid line histogram), followed by Streptavidin-PE (Cat. No. 554061). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The fluorescence histogram showing CD3 expression (or Ig Isotype control staining) was derived from events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
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BD Pharmingen™
CD3E; T3E; TCRE; cd 3; cd-3; cd3; CD3-epsilon; 916
Human (QC Testing)
Mouse IgG2a, κ
Flow cytometry (Routinely Tested), Functional assay, Immunohistochemistry-frozen (Tested During Development)
20 µl
V 5T-CD03.05
AB_395744
Aqueous buffered solution containing BSA, protein stabilizer, and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.

Recommended Assay Procedures

For use of this antibody in immunohistochemical staining of tissue sections, purified anti-human CD3 mAb HIT3a (Cat. No. 550367) is recommended.  For functional assays, our no azide, low endotoxin (NA/LE) mAb HIT3a (Cat. No. 555336) formulation is recommended.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
555338 Rev. 12
Antibody Details
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HIT3a

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).

555338 Rev. 12
Format Details
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Biotin
Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
Biotin
555338 Rev.12
Citations & References
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Development References (7)

  1. Barclay NA, Brown MH, Birkeland ML, et al, ed. The Leukocyte Antigen FactsBook. San Diego, CA: Academic Press; 1997.
  2. Beverley PC, Callard RE. Distinctive functional characteristics of human "T" lymphocytes defined by E rosetting or a monoclonal anti-T cell antibody. Eur J Immunol. 1981; 11(4):329-334. (Biology). View Reference
  3. Guesdon JL, Ternynck T, Avrameas S. The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem. 1979; 27(8):1131-1139. (Biology). View Reference
  4. Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
  5. Lanier LL, Allison JP, Phillips JH. Correlation of cell surface antigen expression on human thymocytes by multi-color flow cytometric analysis: implications for differentiation. J Immunol. 1986; 137(8):2501-2507. (Biology). View Reference
  6. McMichael AJ. A.J. McMichael .. et al., ed. Leucocyte typing III : white cell differentiation antigens. Oxford New York: Oxford University Press; 1987:1-1050.
  7. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
View All (7) View Less
555338 Rev. 12

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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.