Purified anti-human/mouse CD3ε (activation epitope) Antibody

Pricing & Availability
Clone
APA1/1 (See other available formats)
Regulatory Status
RUO
Other Names
CD3e molecule, epsilon (CD3-TCR complex), CD3e antigen, CD3 epsilon
Isotype
Mouse IgG1, κ
APA1_5_Purified_CD3e_Antibody_ICFC_070814
Human peripheral blood lymphocytes were fixed and permeabilized with 70% ethanol, followed by intracellular staining with purified CD3e (clone APA1/1, filled histogram) or mouse IgG1, κ isotype control (open histogram) followed by anti-mouse IgG PE.
  • APA1_5_Purified_CD3e_Antibody_ICFC_070814
    Human peripheral blood lymphocytes were fixed and permeabilized with 70% ethanol, followed by intracellular staining with purified CD3e (clone APA1/1, filled histogram) or mouse IgG1, κ isotype control (open histogram) followed by anti-mouse IgG PE.
  • APA1_5_Purified_CD3e_Antibody_WB_070814
    Western blot analysis of extracts from Jurkat (lane 1) and PBMC untreated (lane 2) and PBMC activated with CD3/CD28(lane 3) cells using anti-CD3ε (activation epitope) antibody (clone APA1/1). β-actin (poly6221) was used as loading control.
  • APA1slash1_PURE_CD3e_activationepitope_Antibody_10142021_IHC-P_1.png
    Human paraffin-embedded tonsil tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris buffer 1X pH 8.5 at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes, blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/mL of purified anti-human CD3ε (clone APA1/1) at 4°C overnight. On the next day, the tissue was washed twice with PBS and stained with goat anti-mouse secondary antibody (clone Poly4053) Alexa Fluor® 594 (red) for two hours at room temperature. The nuclei were counter staining with DAPI (blue). The image was captured with a 10X objective.
  • APA1slash1_PURE_CD3e_activationepitope_Antibody_10142021_IHC-P_2
    Human paraffin-embedded spleen tissue slices were prepared with a standard protocol of deparaffination and rehydration. Antigen retrieval was done with Tris buffer 1X pH 8.5 at 95°C for 40 minutes. Tissue was washed with PBS/ 0.05% Tween20 twice for five minutes, blocked with 5% FBS and 0.2% gelatin for 30 minutes. Then, the tissue was stained with 10 µg/mL of purified anti-human CD3ε (clone APA1/1) and anti-human IgD (clone W18340A) at 4°C overnight. On the next day, the tissue was washed twice with PBS and stained with goat anti-mouse secondary antibody (clone Poly4053) Alexa Fluor® 594 (red) and anti-rat secondary antibody (clone Poly4054) Alexa Fluor® 647 (green) for two hours at room temperature. The nuclei were counter staining with DAPI (blue). The image was captured with a 10X objective.
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362701 50 µg $65.00
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362702 500 µg $258.00
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Description

The CD3ε chain of the CD3-TCR complex is a type I membrane protein that undergoes conformational change following ligation of the TCR-CD3 complex.  This conformational change leads to exposure of the proline rich sequence in CD3ε and results in recruitment of Nck. It is expressed by all mature T cells and forms a heteromeric complex with CD3 α/β or γ/δ and ζ chains. It is important in T cell development, and deficiency of this gene has been associated with severe immunodeficiency.

Technical data sheet

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Purified denatured CD3 proteins from human thymus
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

ICFC - Quality tested
WB, IHC-P - Verified
in vivo cell tracking - Reported in the literature, not verified in house
ICC, IHC, IP - Reported by the developer, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. For Western blotting, the suggested use of this reagent is 1.0 - 4.0 µg per mL. For immunohistochemistry, a concentration range of 5.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone APA1/1 recognizes an activation-dependent intracellular epitope of the CD3ε chain. Additional reported applications (for the relevant formats of this clone) include: immunofluorescent staining of paraformaldehyde fixed sections1, in vivo tracking of T cells 2, and Western blotting3. The LEAF purified format is recommended for in vivo studies. 

Application References

(PubMed link indicates BioLegend citation)
  1. Salmerón A, et al. 1991. J. Immunol. 147:3047. (ICFC, IF)
  2. Risueño RM, et al. 2005. Blood. 106:601. (IF, in vivo tracking, Mouse Reactivity)
  3. Risueño RM, et al. 2008. PLos One. 3:e1747. (WB)
  4. Gil D, et al. 2005. J. Exp. Med. 201:517.
  5. Risueño RM, et al. 2006. Proc. Natl. Acad. Sci. USA. 103:9625.
Product Citations
  1. Srivastava R, et al. 2023. Antib Ther. 253:6. PubMed
  2. Kuai Y, et al. 2020. Theranostics. 5.099305556. PubMed
RRID
AB_2563713 (BioLegend Cat. No. 362701)
AB_2563713 (BioLegend Cat. No. 362702)

Antigen Details

Structure
Type I membrane protein, part of the TCR-CD3 complex. 207 amino acids, predicted molecular weight of 23 kD.
Distribution

Mature T cells.

Function
Important role in T cell development and in linking antigen recognition with intracellular signaling pathways.
Interaction
Interacts with Nck.
Ligand/Receptor
MHC antigen.
Cell Type
T cells
Biology Area
Cell Adhesion, Cell Biology, Immunology, Signal Transduction
Molecular Family
CD Molecules, TCRs
Antigen References

1. Savage PD, et al. 1988. Cytogenet. Cell. Genet. 49:289.
2. DeJarnette JB, et al. 1998. Proc. Natl. Acad. Sci USA. 95:14909.
3. Wang B, et al. 1999. J. Immunol. 162:88.
4. Szymczak AL, et al. 2005. J. Immunol. 175:270.
5. Borroto A, et al. 2014. J. Immunol. 192:2042.

Gene ID
916 View all products for this Gene ID
UniProt
View information about CD3epsilon on UniProt.org
Go To Top Version: 3    Revision Date: 10/14/2021

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Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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