Anti-CD3 antibody (ab16044)
Key features and details
- Rabbit polyclonal to CD3
- Suitable for: IP, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
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- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-CD3 antibody
See all CD3 primary antibodies -
Description
Rabbit polyclonal to CD3 -
Host species
Rabbit -
Tested applications
Suitable for: IP, WBmore details
Unsuitable for: IHC -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human CD3.
Read Abcam's proprietary immunogen policy (Peptide available as ab16206.) -
Positive control
- Recombinant Human CD3 epsilon protein (ab114153) can be used as a positive control in WB. This antibody gave a positive signal in Jurkat whole cells and thymus tissue from Mouse and Rat.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab16044 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 |
|---|---|---|
| IP |
Use a concentration of 5 µg/ml.
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| WB |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
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| Notes |
|---|
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IP
Use a concentration of 5 µg/ml. |
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WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa). |
Target
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Function
The CD3 complex mediates signal transduction. -
Involvement in disease
Defects in CD3D are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)/B(+)/NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. -
Sequence similarities
Contains 1 ITAM domain. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 915 Human
- Entrez Gene: 916 Human
- Entrez Gene: 917 Human
- Entrez Gene: 919 Human
- Entrez Gene: 12500 Mouse
- Entrez Gene: 12501 Mouse
- Entrez Gene: 12502 Mouse
- Entrez Gene: 12503 Mouse
see all -
Alternative names
- 4930549J05Rik antibody
- A430104F18Rik antibody
- AW552088 antibody
see all
Images
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Western blot - Anti-CD3 antibody (ab16044)All lanes : Anti-CD3 antibody (ab16044) at 1 µg/ml
Lane 1 : THP1 whole cell lysate (-ve control)
Lane 2 : Raji whole cell lysate (-ve control)
Lane 3 : Jurkat 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-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 23 kDa
Additional bands at: 17 kDa. We are unsure as to the identity of these extra bands.Lanes 1 - 6: Merged signal (red and green). Green – ab16044 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
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 ab16044 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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Western blot - Anti-CD3 antibody (ab16044)All lanes : Anti-CD3 antibody (ab16044) at 1 µg/ml
Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg
Lane 2 : Thymus (Mouse) Tissue Lysate at 20 µg
Lane 3 : Thymus (Rat) Tissue Lysate at 20 µg
Secondary
All lanes : Anti-Rabbit IgG VHH Single Domain (HRP) (ab191866) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 23 kDa
Additional bands at: 18 kDa, 48 kDa, 62 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 8 minutesThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab16044 overnight at 4°C. Antibody binding was detected using an anti-rabbit IgG VHH single domain antibody conjugated to HRP, and visualised using ECL development solution ab133406.
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Immunoprecipitation - Anti-CD3 antibody (ab16044)CD3 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to CD3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab16044.
Secondary: Clean-Blot IP Detection Reagent (HRP) at 1/500 dilution.
Band: 23kDa; CD3
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (12)
ab16044 has been referenced in 12 publications.
- Hahn L et al. IL-13 as Target to Reduce Cholestasis and Dysbiosis in Abcb4 Knockout Mice. Cells 9:N/A (2020). PubMed: 32846954
- Tian L et al. Calotropin regulates the apoptosis of non-small cell cancer by regulating the cytotoxic T-lymphocyte associated antigen 4-mediated TGF-ß/ERK signaling pathway. Mol Med Rep 17:7683-7691 (2018). PubMed: 29620207
- Carbone ML et al. Topical Plant Polyphenols Prevent Type I Interferon Signaling in the Skin and Suppress Contact Hypersensitivity. Int J Mol Sci 19:N/A (2018). PubMed: 30200670
- Daniels BP et al. Regional astrocyte IFN signaling restricts pathogenesis during neurotropic viral infection. J Clin Invest 127:843-856 (2017). PubMed: 28134626
- Martin JL et al. Inhibition of basal-like breast cancer growth by FTY720 in combination with epidermal growth factor receptor kinase blockade. Breast Cancer Res 19:90 (2017). PubMed: 28778177