Recombinant Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free (ab251607)
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 - BSA and Azide free
- Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-CD3 epsilon antibody [CAL57] - BSA and Azide free
See all CD3 epsilon primary antibodies -
Description
Rabbit monoclonal [CAL57] to CD3 epsilon - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Fr, IP, IHC-P, Flow Cyt (Intra)more 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 (intra): Human peripheral blood mononuclear cells. Mouse splenocytes. IP: EL4.IL-2 whole cell lysate.
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General notes
ab251607 is the carrier-free version of ab237721.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Purification notes
Purity is greater than 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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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251607 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 |
Use at an assay dependent concentration. Predicted molecular weight: 23 kDa.
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IHC-Fr |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 23 kDa. |
IHC-Fr
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
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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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237721).
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
References (0)
ab251607 has not yet been referenced specifically in any publications.