Anti-CD81 antibody [TS81] - BSA and Azide free (ab59477)
Key features and details
- Mouse monoclonal [TS81] to CD81 - BSA and Azide free
- Suitable for: IHC-P, Flow Cyt, WB, ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG2a
Overview
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Product name
Anti-CD81 antibody [TS81] - BSA and Azide free
See all CD81 primary antibodies -
Description
Mouse monoclonal [TS81] to CD81 - BSA and Azide free -
Host species
Mouse -
Specificity
Recognises the TAPA-1 antigen, a 23 kDa (smear) protein,
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Tested applications
Suitable for: IHC-P, Flow Cyt, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Tissue, cells or virus corresponding to CD81. Jurkat cell line
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Positive control
- This antibody gave a positive result in IHC in the following FFPE tissue: Human normal lung. Flow Cyt: HAP1-WT cells. WB: Raji (Human Burkitt's lymphoma cell line) whole cell lysate
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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: 7.30
Constituent: 100% PBS
Sterile-filtered through 0.22 µm. Carrier and preservative free -
Carrier free
Yes -
Concentration information loading...
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Purity
Ion Exchange Chromatography -
Purification notes
ab59477 is sterile-filtered through 0.22 µm and treated to remove endotoxins. -
Clonality
Monoclonal -
Clone number
TS81 -
Myeloma
x63-Ag8.653 -
Isotype
IgG2a -
Light chain type
kappa -
Research areas
Associated products
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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 ab59477 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 |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt |
Use 0.5µg for 106 cells.
ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Use under non reducing condition. Predicted molecular weight: 25 kDa.
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ICC/IF |
Use a concentration of 1 µg/ml.
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Notes |
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IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt
Use 0.5µg for 106 cells. ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Use under non reducing condition. Predicted molecular weight: 25 kDa. |
ICC/IF
Use a concentration of 1 µg/ml. |
Target
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Function
May play an important role in the regulation of lymphoma cell growth. Interacts with a 16-kDa Leu-13 protein to form a complex possibly involved in signal transduction. May acts a the viral receptor for HCV. -
Tissue specificity
Hematolymphoid, neuroectodermal and mesenchymal tumor cell lines. -
Involvement in disease
Defects in CD81 are the cause of immunodeficiency common variable type 6 (CVID6) [MIM:613496]; also called antibody deficiency due to CD81 defect. CVID6 is a primary immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent bacterial infections and an inability to mount an antibody response to antigen. The defect results from a failure of B-cell differentiation and impaired secretion of immunoglobulins; the numbers of circulating B cells is usually in the normal range, but can be low. -
Sequence similarities
Belongs to the tetraspanin (TM4SF) family. -
Post-translational
modificationsNot glycosylated. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 975 Human
- Omim: 186845 Human
- SwissProt: P60033 Human
- Unigene: 54457 Human
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Alternative names
- 26 kDa cell surface protein TAPA 1 antibody
- 26 kDa cell surface protein TAPA-1 antibody
- 26 kDa cell surface protein TAPA1 antibody
see all
Images
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All lanes : Anti-CD81 antibody [TS81] - BSA and Azide free (ab59477)
Lane 1 : Positive cell line Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 25 µg
Lane 2 : Positive cell line total protein in non-reducing conditions at 50 µg
Lane 3 : Negative cell line U266 at 25 µg
Lane 4 : Negative cell line total protein in non-reducing conditions at 50 µg
Predicted band size: 25 kDaWestern Blot under non-reducing conditions
Detection: chemiluminescence
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ICC/IF image of CD81 stained Hek293 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab59477, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
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Overlay histogram showing Jurkat cells stained with ab59477 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab59477, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
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Overlay histogram showing HAP1 wildtype (green line) and HAP1-CD81 knockout cells (red line) stained with ab59477. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab59477, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) presorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.
A mouse IgG1 isotype control antibody (ab170190) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CD81 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
This antibody can also be used in HAP1 cells fixed with 80% methanol (5 min), , permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions. -
IHC image of CD81 staining in Human normal lung formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab59477, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then 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.
Protocols
Datasheets and documents
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Datasheet download
References (18)
ab59477 has been referenced in 18 publications.
- Xie H et al. Exosome-transmitted circVMP1 facilitates the progression and cisplatin resistance of non-small cell lung cancer by targeting miR-524-5p-METTL3/SOX2 axis. Drug Deliv 29:1257-1271 (2022). WB ; Human . PubMed: 35467477
- Han Z et al. Matrix-assisted laser desorption ionization mass spectrometry profiling of plasma exosomes evaluates osteosarcoma metastasis. iScience 24:102906 (2021). PubMed: 34401680
- Zhang X et al. Proteomic analysis of the exosomes secreted from Ctenopharyngodon idellus kidney cells infected with grass carp reovirus reveals their involvement in the cellular responses to viral infection. Fish Physiol Biochem 47:857-867 (2021). PubMed: 33745109
- Berzaghi R et al. Secretion rates and protein composition of extracellular vesicles released by cancer-associated fibroblasts after radiation. J Radiat Res 62:401-413 (2021). PubMed: 33899109
- Chen K et al. Inhibition of Circulating Exosomal miRNA-20b-5p Accelerates Diabetic Wound Repair. Int J Nanomedicine 16:371-381 (2021). PubMed: 33469291