Recombinant Anti-CD3 zeta antibody [BL-336-1B2] - BSA free (ab272071)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [BL-336-1B2] to CD3 zeta - BSA free
- Suitable for: WB, IHC-P, IP
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD3 zeta antibody [BL-336-1B2] - BSA free
See all CD3 zeta primary antibodies -
Description
Rabbit monoclonal [BL-336-1B2] to CD3 zeta - BSA free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human CD3 zeta aa 150-164 (C terminal). The exact sequence is proprietary. NP_932170.1 and Gene ID 919.
Database link: P20963 -
Positive control
- WB: K562, Jurkat, HeLa, SR, HEK293T, HepG2 and MOLT4 whole cell lysate. IHC-P: Human tonsil tissue, human lung cancer tissue. IP: Jurkat whole cell lysate.
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General notes
ab272071 is the BSA-free version of ab243874.
This product is sold under License from Bethyl Laboratories, Inc.
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: 8.20
Preservative: 0.09% Sodium azide
Constituent: 98% Borate buffered saline -
Concentration information loading...
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Purification notes
Recombinant antibody was purified from cell culture supernatant. -
Clonality
Monoclonal -
Clone number
BL-336-1B2 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab272071 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.
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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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IP |
Use at an assay dependent concentration.
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Notes |
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WB
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. |
IP
Use at an assay dependent concentration. |
Target
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Function
Probable role in assembly and expression of the TCR complex as well as signal transduction upon antigen triggering. -
Involvement in disease
Defects in CD247 are the cause of immunodeficiency due to defect in CD3-zeta (CD3ZID) [MIM:610163]. An immunological deficiency characterized by T-cells impaired immune response to alloantigens, tetanus toxoid and mitogens. -
Sequence similarities
Belongs to the CD3Z/FCER1G family.
Contains 3 ITAM domains. -
Domain
The ITAM domains mediate interaction with SHB. -
Post-translational
modificationsPhosphorylated on Tyr residues after T-cell receptor triggering. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 919 Human
- Omim: 186780 Human
- SwissProt: P20963 Human
- Unigene: 156445 Human
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Alternative names
- 4930549J05Rik antibody
- A430104F18Rik antibody
- AW552088 antibody
see all
Images
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All lanes : Anti-CD3 zeta antibody [BL-336-1B2] (ab243874) at 1/1000 dilution
Lane 1 : Wild-type Jurkat cell lysate
Lane 2 : CD247 knockout Jurkat cell lysate
Lane 3 : MOLT-4 cell lysate
Lane 4 : K562 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Observed band size: 16 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-CD3 zeta antibody [BL-336-1B2] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab243874 was shown to bind specifically to CD3 zeta. A band was observed at 16 kDa in wild-type Jurkat cell lysates with no signal observed at this size in CD247 knockout cell line ab273856 (knockout cell lysate ab273810). To generate this image, wild-type and CD247 knockout Jurkat cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing Borate buffered saline, BSA, glycerol and sodium azide (ab243874).
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All lanes : Anti-CD3 zeta antibody [BL-336-1B2] (ab243874) at 1/1000 dilution
Lane 1 : K562 whole cell lysate
Lane 2 : Jurkat whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : SR whole cell lysate
Lane 5 : HEK293T whole cell lysate
Lane 6 : HepG2 whole cell lysate
Lane 7 : MOLT4 whole cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG
Exposure time: 10 secondsLysates prepared using NETN lysis buffer.
This data was developed using the same antibody clone in a different buffer formulation containing Borate buffered saline, BSA, glycerol and sodium azide (ab243874).
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD247/CD3Z with ab243874. HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody, with DAB as substrate.
This data was developed using the same antibody clone in a different buffer formulation containing Borate buffered saline, BSA, glycerol and sodium azide (ab243874).
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CD247/CD3Z was immunoprecipitated from 1mg of Jurkat whole cell lysate with ab243874 at 20 µl/mg lysate. Western blot was performed from the immunoprecipitate using ab264158 at a 1/1000 dilution.
Lane 1: ab243874 IP in Jurkat whole cell lysate.
Lane 2: Control IgG.Detection: Chemiluminescence.
Lysate prepared using NETN lysis buffer.
This data was developed using the same antibody clone in a different buffer formulation containing Borate buffered saline, BSA, glycerol and sodium azide (ab243874).
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue, labeling CD247/CD3Z with ab243874. HRP-conjugated goat anti-rabbit IgG used as the secondary antibody. DAPI counterstain.
This data was developed using the same antibody clone in a different buffer formulation containing Borate buffered saline, BSA, glycerol and sodium azide (ab243874).
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Immunohistochemical analysis of formalin-fixed, paraffin-embedded human lung cancer tissue labeling CD247/CD3Z with ab243874. HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody, with DAB as the substrate.
This data was developed using the same antibody clone in a different buffer formulation containing Borate buffered saline, BSA, glycerol and sodium azide (ab243874).
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)
ab272071 has not yet been referenced specifically in any publications.