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Anti-CD3 antibody [SP162] is a rabbit recombinant monoclonal antibody that is used to detect CD3 in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse samples.

- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 50 publications


Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP162] (AB135372), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-CD3 antibody [SP162] (AB135372), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (AB135372), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-CD3 antibody [SP162] (AB135372), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (AB135372), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.6
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PFlow Cyt (Intra)WBICC/IF
Human
Tested
Tested
Tested
Tested
Mouse
Expected
Tested
Expected
Tested
Rat
Predicted
Predicted
Predicted
Predicted
Chicken
Predicted
Predicted
Predicted
Predicted
Cow
Predicted
Predicted
Predicted
Predicted
Dog
Predicted
Predicted
Predicted
Predicted
Pig
Predicted
Predicted
Predicted
Predicted
Rabbit
Predicted
Predicted
Predicted
Predicted

Tested
Tested

Species
Human
Dilution info
1/150
Notes

Incubate with primary antibody for 10 minutes at room temperature.

Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Rat, Rabbit, Chicken, Cow, Dog, Pig
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/50 - 1/200
Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Species
Human
Dilution info
1/50 - 1/200
Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Predicted
Predicted

Species
Rat, Rabbit, Chicken, Cow, Dog, Pig
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/150
Notes

Incubate for 1 hour at room temperature.

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Predicted
Predicted

Species
Rat, Rabbit, Chicken, Cow, Dog, Pig
Dilution info
-
Notes

-

Tested
Tested

Species
Mouse
Dilution info
1/10
Notes

-

Species
Human
Dilution info
1/10
Notes

-

Predicted
Predicted

Species
Rat, Rabbit, Chicken, Cow, Dog, Pig
Dilution info
-
Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

1 product for Alternative Version

Target data

Function

Part of the TCR-CD3 complex present on T-lymphocyte cell surface that plays an essential role in adaptive immune response. When antigen presenting cells (APCs) activate T-cell receptor (TCR), TCR-mediated signals are transmitted across the cell membrane by the CD3 chains CD3D, CD3E, CD3G and CD3Z. All CD3 chains contain immunoreceptor tyrosine-based activation motifs (ITAMs) in their cytoplasmic domain. Upon TCR engagement, these motifs become phosphorylated by Src family protein tyrosine kinases LCK and FYN, resulting in the activation of downstream signaling pathways (PubMed:2470098). In addition of this role of signal transduction in T-cell activation, CD3E plays an essential role in correct T-cell development. Initiates the TCR-CD3 complex assembly by forming the two heterodimers CD3D/CD3E and CD3G/CD3E. Participates also in internalization and cell surface down-regulation of TCR-CD3 complexes via endocytosis sequences present in CD3E cytosolic region (PubMed:10384095, PubMed:26507128). In addition to its role as a TCR coreceptor, it serves as a receptor for ITPRIPL1. Ligand recognition inhibits T-cell activation by promoting interaction with NCK1, which prevents CD3E-ZAP70 interaction and blocks the ERK-NFkB signaling cascade and calcium influx (PubMed:38614099).

Alternative names

CD3e, T3E, CD3E, T-cell surface glycoprotein CD3 epsilon chain, T-cell surface antigen T3/Leu-4 epsilon chain

Recommended products

Anti-CD3 antibody [SP162] is a rabbit recombinant monoclonal antibody that is used to detect CD3 in Flow cytometry (Intra), ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse samples.

- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Cited in over 50 publications

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
SP162
Purification technique
Affinity purification Protein A/G
Concentration
Loading...
Purification notes

Purified from TCS by protein A/G.

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
Do Not Freeze

Notes

Anti-CD3 antibody [SP162] (ab135372) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), ICC/IF, IHC-P and WB.

Abcam's high quality manufacturing and validation processes ensure Anti-CD3 antibody [SP162] (ab135372) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

Anti-CD3 antibody [SP162] (ab135372) has 4 independent reviews from customers.

Anti-CD3 antibody [SP162] (ab135372) specifically detects CD3 (UniProt ID: P07766; Molecular weight: 21kDa) and is sold in 100 µL, 500 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone SP162 - Anti-CD3 antibody [SP162] - BSA and Azide free ab245731.

Antibody clone SP162 is also available pre-conjugated to a variety of labels for your convenience - APC, HRP, Alexa Fluor® 488, Alexa Fluor® 647, Alexa Fluor® 555, Alkaline Phosphatase (APC Anti-CD3 antibody [SP162] ab305633, HRP Anti-CD3 antibody [SP162] ab305634, Alexa Fluor® 488 Anti-CD3 antibody [SP162] ab307134, Alexa Fluor® 647 Anti-CD3 antibody [SP162] ab307265, Alexa Fluor® 555 Anti-CD3 antibody [SP162] ab307335, Alkaline Phosphatase Anti-CD3 antibody [SP162] ab308972).

Anti-CD3 antibody [SP162] (ab135372) is a clone from the portfolio of Spring Bioscience (Roche) SP clones which have been optimised for immunohistochemistry (IHC).

CD3 is a crucial T cell co-receptor involved in activating both cytotoxic (CD8+) and helper (CD4+) T cells, playing a pivotal role in immune response. In immuno-oncology, targeting CD3 with therapeutic antibodies can enhance T cell-mediated anti-tumor activity, making it a valuable strategy in cancer treatment.

What are the advantages of a recombinant monoclonal antibody?

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

8 product images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP162] (ab135372)

    Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD3 with ab135372 at a 1/150 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD3 antibody [SP162] (ab135372)

    Immunocytochemistry/ Immunofluorescence analysis of Jurkat (human T cell leukemia T lymphocyte) cells labeling CD3 with purified ab135372 at 1/10 (10 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (ab135372)

    Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CD3 with purified ab135372 at 1/200 dilution (0.60 μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

  • Immunocytochemistry/ Immunofluorescence - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-CD3 antibody [SP162] (ab135372)

    Immunocytochemistry/ Immunofluorescence analysis of EL4.IL-2 (mouse lymphoma T lymphocyte) cells labeling CD3 with purified ab135372 at 1/10 (10 μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1/1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (ab135372)

    Flow Cytometry analysis of Mouse Splenocyte cells labeling CD3 with purified ab135372 at 1/20 dilution (5.95 μg/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Black. Unlabeled control - Unlabelled cells / blue.

  • Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-CD3 antibody [SP162] (ab135372)

    Flow cytometric analysis of rabbit anti-CD3 (SP162) antibody ab135372 (1/150) in Jurkat (Human T cell leukemia cell line from peripheral blood) cells (green) compared to negative control of rabbit IgG (blue).

  • Western blot - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Western blot - Anti-CD3 antibody [SP162] (ab135372)

    All lanes: Western blot - Anti-CD3 antibody [SP162] (ab135372) at 1/150 dilution

    All lanes: Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate

    Predicted band size: 113 kDa, 13 kDa, 14 kDa, 16 kDa, 19 kDa, 30 kDa, 40 kDa, 41 kDa, 51 kDa, 53 kDa, 58 kDa, 59 kDa, 60 kDa, 67 kDa, 68 kDa

    Observed band size: 14 kDa, 60 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP162] (ab135372), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP162] (ab135372)

    Immunohistochemical analysis of formalin fixed paraffin embedded Human tonsil labelling CD3 with ab135372 at a dilution of 1/300. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

    ab135372 anti CD3 antibody SP162 was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

    Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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