Recombinant Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22667-12] to CD3 epsilon
- Suitable for: WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse
Related conjugates and formulations
Overview
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Product name
Anti-CD3 epsilon antibody [EPR22667-12]
See all CD3 epsilon primary antibodies -
Description
Rabbit monoclonal [EPR22667-12] to CD3 epsilon -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, ICC/IF, IPmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: EL4.IL2 whole cell lysate; mouse thymus lysate. IP: EL4.IL2 whole cell lysate. IHC-P: Mouse spleen and lung tissue. ICC/IF: EL4.IL2 cells.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22667-12 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab231775 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/1000. Detects a band of approximately 18, 22 kDa (predicted molecular weight: 21 kDa).
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IHC-P |
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/100.
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IP |
1/30.
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Notes |
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WB
1/1000. Detects a band of approximately 18, 22 kDa (predicted molecular weight: 21 kDa). |
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
1/100. |
IP
1/30. |
Target
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Function
The CD3 complex mediates signal transduction. -
Sequence similarities
Contains 1 Ig-like (immunoglobulin-like) domain.
Contains 1 ITAM domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 12501 Mouse
- SwissProt: P22646 Mouse
- Unigene: 210361 Mouse
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Alternative names
- CD3 epsilon antibody
- CD3e antibody
- CD3e antigen antibody
see all
Images
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EL4.IL2 (mouse lymphoma T lymphocyte cell line) cells labeling CD3 epsilon with ab231775 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining in EL4.IL2 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 epsilon with ab231775 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the mouse spleen (PMID: 18648532) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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CD3 epsilon was immunoprecipitated from 0.35 mg of EL4.IL2 (mouse lymphoma T lymphocyte cell line) whole cell lysate with ab231775 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab231775 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as the secondary antibody at 1/5000 dilution.
Lane 1: EL4.IL2 whole cell lysate 10 μg (Input).
Lane 2: ab231775 IP in EL4.IL2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab231775 in EL4.IL2 whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds. -
Anti-CD3 epsilon antibody [EPR22667-12] (ab231775) at 1/1000 dilution + EL4.IL2 (mouse lymphoma T lymphocyte cell line) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 21 kDa
Observed band size: 22 kDa why is the actual band size different from the predicted?
Exposure time: 158 secondsBlocking and dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [EPR22667-12] (ab231775)
Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling CD3 epsilon with ab231775 at 1/5000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining on the T lymphocytes on mouse lung (PMID: 22888136) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.
Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Anti-CD3 epsilon antibody [EPR22667-12] (ab231775) at 1/1000 dilution + Mouse thymus lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 21 kDa
Observed band size: 18,22 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking and dilution buffer: 5% NFDM/TBST.
The expression profile observed is consistent with what has been described in the literature (PMID: 1387664).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab231775 has been referenced in 1 publication.
- Yi M et al. Combine and conquer: manganese synergizing anti-TGF-ß/PD-L1 bispecific antibody YM101 to overcome immunotherapy resistance in non-inflamed cancers. J Hematol Oncol 14:146 (2021). PubMed: 34526097