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Mouse CD3ε Activating (145-2C11) Hamster mAb (Low Endotoxin, Azide-free)
Primary Antibodies
Monoclonal Antibody

Mouse CD3ε Activating (145-2C11) Hamster mAb (Low Endotoxin, Azide-free) #35425

Citations (1)
To Purchase # 35425
Cat. # Size Qty. Price
100 µg

Supporting Data

MW (kDa)
Source/Isotype Hamster (armenian) IgG

Application Key:

  • WB-Western Blot
  • IP-Immunoprecipitation
  • IHC-Immunohistochemistry
  • ChIP-Chromatin Immunoprecipitation
  • C&T-CUT&Tag
  • DB-Dot Blot
  • IF-Immunofluorescence
  • F-Flow Cytometry

Species Cross-Reactivity Key:

  • H-Human
  • M-Mouse
  • R-Rat
  • Hm-Hamster
  • Mk-Monkey
  • Vir-Virus
  • Mi-Mink
  • C-Chicken
  • Dm-D. melanogaster
  • X-Xenopus
  • Z-Zebrafish
  • B-Bovine
  • Dg-Dog
  • Pg-Pig
  • Sc-S. cerevisiae
  • Ce-C. elegans
  • Hr-Horse
  • GP-Guinea Pig
  • Rab-Rabbit
  • All-All Species Expected

Product Description

Endotoxin Less than or equal to 0.01 EU/μg, as determined by the LaL assay.


Supplied in 10 mM NaH2PO4, 150 mM NaCl, pH 7.2, Low Endotoxin, Azide-free.


Stable for 12 months when stored at 4°C. Do not aliquot the antibody.



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A. Solutions and Reagents

  1. Mouse CD3ε Activating (145-2C11) Hamster mAb (Low Endotoxin, Azide-free) #35425
  2. Mouse CD28 Activating (37.51) Hamster mAb (Low Endotoxin, Azide-free) #34596

B. Stimulation Protocol: Plate-based

  • Prepare a 10 µg/ml solution of CD3ε Activating (145-2C11) Hamster mAb (Low Endotoxin, Azide-free) #35425 in sterile PBS. Prepare a sufficient volume to cover the bottom of all the wells to be used for the stimulated T cell condition (approximately 50 µl for 96-well plates, 1 ml for 6-well plates).
  • Add the diluted CD3ε (145-2C11) antibody solution to the wells chosen for treatment. Add equivalent volume of sterile PBS to the wells chosen as controls for treatment.
  • Cover the plate and incubate at 37 °C for a minimum of 2 hrs. The plate may be allowed to incubate overnight if desired.
  • Prepare cells as needed to generate a cell suspension of roughly 1 x 106 cells/ml in a sufficient volume of complete media (100 µl per well for 96-well plates, 2 ml for 6-well plates). Optimal cell density may be determined through experimentation with cells of interest. For T cell expansion, add Mouse IL-2 Recombinant Protein #68966 to the cell suspension to yield a final concentration of 20 ng/ml.
  • Aspirate the CD3ε (145-2C11) antibody solution from the wells, and rinse wells 2x with excess sterile PBS to remove unbound antibody.
  • Add cell suspension to all wells, at volumes specified in step 4 or as determined by experimentation.
  • Add Mouse CD28 Activating (37.51) Hamster mAb (Low Endotoxin, Azide-free) #34596 to yield a concentration of 2 µg/ml. Pre-dilute CD28 (37.51) antibody in complete media if needed. CD28 antibody may also be added to a portion of control wells if desired.
  • Incubate cells for desired time in a humidified incubator (37 °C, 5% CO2). Add fresh media with IL-2 as needed for T cell expansion.
  • Harvest cells and proceed with immunostaining or analysis as desired.

Protocol Id: 2865

Specificity / Sensitivity

Mouse CD3ε Activating (145-2C11) Hamster mAb (Low Endotoxin, Azide-free) recognizes endogenous levels of CD3ε protein.

Species Reactivity:


Source / Purification

Monoclonal antibody is produced against mouse BM10-37 cytotoxic T cells and purified via affinity chromatography.


When T cells encounter antigens via the T cell receptor (TCR), information about the quantity and quality of antigens is relayed to the intracellular signal transduction machinery (1). This activation process depends mainly on CD3 (Cluster of Differentiation 3), a multiunit protein complex that directly associates with the TCR. CD3 is composed of four polypeptides: ζ, γ, ε, and δ. Each of these polypeptides contains at least one immunoreceptor tyrosine-based activation motif (ITAM) (2). Engagement of the TCR complex with foreign antigens induces tyrosine phosphorylation in the ITAM motifs and phosphorylated ITAMs function as docking sites for signaling molecules such as ZAP-70 and the p85 subunit of PI-3 kinase (3,4). TCR ligation also induces a conformational change in CD3ε, such that a proline region is exposed and then associates with the adaptor protein Nck (5).

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For Research Use Only. Not for Use in Diagnostic Procedures.
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