Recombinant Anti-CD3 antibody [CD3-12] - BSA and Azide free (ab255972)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rat monoclonal [CD3-12] to CD3 - BSA and Azide free
- Suitable for: IHC-P, Flow Cyt (Intra), WB
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-CD3 antibody [CD3-12] - BSA and Azide free
See all CD3 primary antibodies -
Description
Rat monoclonal [CD3-12] to CD3 - BSA and Azide free -
Host species
Rat -
Tested applications
Suitable for: IHC-P, Flow Cyt (Intra), WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human tonsil tissue. Mouse spleen and lymph node tissue; Rat spleen tissue. Flow Cyt: Human peripheral blood lymphocytes. WB: Jurkat, MOLT-4 and EL4 cell lysates; Human, mouse and rat thymus tissue lysates.
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General notes
ab255972 is the carrier-free version of ab11089.
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
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.
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
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 -
Clonality
Monoclonal -
Clone number
CD3-12 -
Isotype
IgG1 -
Research areas
Associated products
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Alternative Versions
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab255972 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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IHC-P | (2) |
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/400.
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WB |
1/1000. Predicted molecular weight: 19 kDa.
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Notes |
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IHC-P
1/200. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/400. |
WB
1/1000. Predicted molecular weight: 19 kDa. |
Images
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
Flow cytometric analysis of 4% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cells (PBMC) labeling CD3 with ab11089 at 1/400 dilution (red) compared with a Mouse IgG, monoclonal Isotype Control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Mouse IgG ab150157 (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1 µg/ml
Lane 1 : THP-1 (Human monocytic leukemia cell line) whole cell lysate (negative control)
Lane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate (negative control)
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 4 : Human thymus tissue lysate
Lane 5 : Mouse thymus tissue lysate
Lane 6 : Rat thymus tissue lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat anti-Rat at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 19 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab11089 overnight at 4°C. Antibody binding was detected using Goat anti-Rat secondary at a 1:10000 dilution for 1hr at room temperature and then imaged.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CB3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat spleen tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse spleen tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
IHC image of CD3 staining in a formalin fixed, paraffin embedded human tonsil tissue. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
IHC image of CD3 staining in a formalin fixed, paraffin embedded normal mouse lymph node tissue. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution
Lane 1 : Mouse thymus tissue lysate
Lane 2 : Rat thymus tissue lysate
Lane 3 : Human thymus tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution
Predicted band size: 19 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure: Lanes 1/3: 5 secs; Lanes 2: 3 secs.
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All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte)
Lane 2 : MOLT-4 (human lymphoblastic leukemia T lymphoblast)
Lane 3 : EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution
Predicted band size: 19 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab11089).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 5 secs.
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)
ab255972 has not yet been referenced specifically in any publications.