Recombinant Anti-CD3 epsilon antibody [CAL57] (ab237721)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CAL57] to CD3 epsilon
- Suitable for: WB, IHC-Fr, Flow Cyt, IP, IHC-P
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-CD3 epsilon antibody [CAL57]
See all CD3 epsilon primary antibodies -
Description
Rabbit monoclonal [CAL57] to CD3 epsilon -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Fr, Flow Cyt, IP, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Mouse CD3 epsilon aa 135-189. The exact sequence is proprietary.
Database link: P22646 -
Positive control
- IHC-P: Mouse and rat spleen tissue. Human tonsil and colon tissue. WB: EL4.IL-2 and Jurkat whole cell lysate. Mouse lymph node lysate. Rat thymus and spleen lysate. IHC-Fr: Mouse thymus tissue. Flow Cyt: Human peripheral blood mononuclear cells. Mouse splenocytes. IP: EL4.IL-2 whole cell lysate.
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: 7.50
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purity >99%. -
Clonality
Monoclonal -
Clone number
CAL57 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab237721 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. Predicted molecular weight: 23 kDa.
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IHC-Fr |
1/500.
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Flow Cyt |
1/500.
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IP |
1/30.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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WB
1/1000. Predicted molecular weight: 23 kDa. |
IHC-Fr
1/500. |
Flow Cyt
1/500. |
IP
1/30. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
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: 916 Human
- Entrez Gene: 12501 Mouse
- Entrez Gene: 315609 Rat
- Omim: 186830 Human
- SwissProt: P07766 Human
- SwissProt: P22646 Mouse
- Unigene: 3003 Human
- Unigene: 210361 Mouse
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Alternative names
- CD3 epsilon antibody
- CD3e antibody
- CD3e antigen antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] (ab237721)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the human tonsil. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] (ab237721)
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the human colon. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse thymus tissue labeling CD3 epsilon with ab237721 at 1/500 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on T cell surface (PMID: 8490660) is observed. Counterstained with DAPI (blue). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1,000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).
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All lanes : Anti-CD3 epsilon antibody [CAL57] (ab237721) at 1/1000 dilution
Lane 1 : EL4.IL-2 (Mouse lymphoma T lymphocyte) whole cell lysate
Lane 2 : Mouse lymph node lysate
Lane 3 : Rat thymus lysate
Lane 4 : Rat spleen lysate
Lane 5 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Rabbit monoclonal [CAL57] to CD3 epsilon (ab237721) at 1/100000 dilution
Predicted band size: 23 kDa
Exposure time: 3 minutesThe expression pattern is consistent with what described in the literature. (PMID: 19542373).
Blocking/Dilution buffer: 5% NFDM/TBST.
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Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse splenocytes labeling CD3 epsilon with ab237721 at 1/500 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab237721 (Right). They recognize the same cell population.
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Flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cell (PBMC) labeling CD3 epsilon with ab237721 at 1/500 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor® 488, ab150097), at 1/5000 dilution was used as the secondary antibody.
Cells were surface stained with anti-CD3 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab237721 (Right). They recognize the same cell population.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] (ab237721)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the rat spleen. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] (ab237721)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 epsilon with ab237721 at 1/2000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the T lymphocytes in the mouse spleen. The section was incubated with ab237721 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
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CD3 epsilon was immunoprecipitated from 0.35 mg EL4.IL2 (Mouse lymphoma T lymphocyte) whole cell lysate with ab237721 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237721 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
Lane 1: EL4.IL2 whole cell lysate 10 µg (Input).
Lane 2: ab237721 IP in EL4.IL2 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237721 in EL4.IL2 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 5 seconds. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 epsilon antibody [CAL57] (ab237721)
Formalin-fixed, paraffin-embedded mouse spleen tissue stained for CD3 epsilon using ab237721 at 0.5 µg/ml in immunohistochemical analysis.
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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SDS download
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Datasheet download
References (0)
ab237721 has not yet been referenced specifically in any publications.