CD81 antibody | Eat2
Hamster anti Mouse CD81 antibody, clone EAT2 (MCA1846) used to evaluate CD81 expression in rat growth cone particles (GCPs) by western blotting.
Image caption:
Presence of APP in GC adhesions.
B. Isolation of GC adhesions on laminin. Rat GCPs plated on laminin were extracted with Brij98 to yield the unattached soluble fraction (see cartoon, green and blue; Sol). The remaining adherent structures (Adh, red) were recovered with SDS. The fractions were analyzed by western blot (equal fractional protein amounts loaded). Samples not extracted with Brij98 served as controls. The adhesion markers Itgb1, CD81 and FAK were enriched in the adherent fraction compared to its soluble counterpart, and significant amounts of APP and Dab1 (upper band) were detected.
From: Sosa LJ, Bergman J, Estrada-Bernal A, Glorioso TJ, Kittelson JM, Pfenninger KH.
Amyloid precursor protein is an autonomous growth cone adhesion molecule engaged in contact guidance.
PLoS One. 2013 May 14;8(5):e64521.
doi: 10.1371/journal.pone.0064521.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Hamster anti Mouse CD81 antibody, clone EAT2 (MCA1846) used to evaluate CD81 expression in rat growth cone particles (GCPs) by western blotting.
Image caption:
Presence of APP in GC adhesions.
C. Co-immunoprecipitation of APP with Itga3 and CD81. The blots on the left show the enrichment of APP in the precipitates (N- or C-terminus-specific antibodies), while those on the right show precipitated Itgb1 (β1) and CD81.
From: Sosa LJ, Bergman J, Estrada-Bernal A, Glorioso TJ, Kittelson JM, Pfenninger KH.
Amyloid precursor protein is an autonomous growth cone adhesion molecule engaged in contact guidance.
PLoS One. 2013 May 14;8(5):e64521.
doi: 10.1371/journal.pone.0064521.
This image is from an open access article distributed under terms of a Creative Commons Attribution License.
Hamster anti mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in RAW264.7 lysates by western blotting.
Image caption:Statins transfer CD14 from lipid rafts into CD9-enriched microdomains. (A) RAW264.7 cells were stimulated with 0.1 µg/ml LPS and, after the indicated times, the cells were lysed and protein levels were examined by immunoblotting. Anti-actin blots show that comparable amounts of protein were loaded in each lane. (B) RAW264.7 cells were untreated (-) or cultured for 24 h in the absence (-) or presence of 5 μM fluvastatin (Fluv) or simvastatin (Simv) (+) and stimulated for 2 h with 1 μg/ml LPS (+). Proteins in whole-cell lysate (WCL) and CD14 protein in immunoprecipitates (IP) with anti-TLR4 Ab were immunoblotted (IB). (C) RAW264.7 cells were treated as in B. Lysates of untreated (C, control) cultures or LPS-stimulated cultures in the absence (L) or presence of fluvastatin (FL) or simvastatin (SL) were fractionated by sucrose density gradients, and protein distributions were visualized by immunoblotting. The intensities of blots were quantified by densitometry, and percentages of density units of light membrane (LM) fractions are displayed to the right of the blots. Data shown are from one representative of three similar experiments. (D) Immunoblots of CD9 and CD81 proteins in whole-cell lysates and in immunoprecipitates with control IgG or anti-CD14 mAb. (E) Immunoblots of CD9 and CD81 proteins in whole-cell lysates and in immunoprecipitates with control IgG or anti-CD14 mAb from pooled LM fractions (4 and 5) and dense (D) fractions (9 and 10). In the presence of statins, more CD14/CD9 complexes were formed in dense fractions (arrowheads).
From: Jin Y, Tachibana I, Takeda Y, He P, Kang S, et al. (2013)
Statins Decrease Lung Inflammation in Mice by Upregulating Tetraspanin CD9 in Macrophages.
PLoS ONE 8(9): e73706.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Hamster anti mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in RAW264.7 lysates by western blotting.
Image caption:
Screening of a drug library for agents that upregulate CD9 or CD81 in RAW264.7 macrophages. (A) RAW264.7 cells were cultured for 24 h in the absence (V, vehicle alone) and presence of each drug (10 μM). The cells were lysed, and levels of CD9 and CD81 were examined by immunoblotting. Blots of results with fluvastatin (Fluv) and simvastatin (Simv) are shown. Anti-actin blots show that comparable amounts of protein were loaded in each lane. (B) After testing 1,165 drugs, levels of CD9 and CD81 relative to actin were quantified by densitometry. Fold changes of the expression levels compared with vehicle alone were calculated and plotted. Drugs that increased the level of either CD9 or CD81 more than 1.5-fold compared with vehicle alone were regarded as positive. Correlation between fold changes in CD9 and CD81 levels was analyzed using Pearson's correlation coefficient. (C) RAW264.7 cells were cultured in the absence (V) or presence of multiple statins (10 µM) and levels of CD9 and CD81 were examined by immunoblotting. The statins are arranged in order of decreasing lipophilicity. Ceri, cerivastatin; Simv, simvastatin; Fluv, fluvastatin; Ator, atorvastatin; Rosu, rosuvastatin; Prav, pravastatin. (D) RAW264.7 cells were cultured in the absence (shaded histograms) or presence (10 μM) of fluvastatin (open red histograms) and simvastatin (open blue histograms). Surface levels of CD9, CD63, CD81, and the integrin β1 subunit were analyzed by flow cytometry.
From: Jin Y, Tachibana I, Takeda Y, He P, Kang S, et al. (2013)
Statins Decrease Lung Inflammation in Mice by Upregulating Tetraspanin CD9 in Macrophages.
PLoS ONE 8(9): e73706.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Hamster anti mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in RAW264.7 lysates by western blotting.
Image caption:
Fluvastatin and simvastatin increase CD9 and CD81 levels in RAW264.7 cells. (A) RAW264.7 cells were cultured for 24 h in the absence or presence of increasing concentrations of fluvastatin (Fluv) or simvastatin (Simv). The cells were lysed, and levels of CD9, CD63, and CD81 were examined by immunoblotting. Anti-actin blots show that comparable amounts of protein were loaded in each lane. (B) RAW264.7 cells were untreated (-) or cultured in the absence or presence of increasing concentrations of fluvastatin or simvastatin and stimulated for 24 h with 0.1 μg/ml LPS (+). Levels of CD9, CD63, and CD81 were examined by immunoblotting. Note that LPS downregulates CD9 and CD81 in the absence of statins (arrowheads). (C) RAW264.7 cells were cultured in the absence (-) or presence of 3 µM fluvastatin (+), and unstimulated (-) or stimulated for 24 h with 1 μg/ml LPS (+). mRNA levels of CD9 and CD81 were examined by reverse transcription PCR. GAPDH is an internal loading control. (D) RAW264.7 cells were cultured in the absence or presence of fluvastatin, and unstimulated or stimulated with LPS. Control (Cont) was an untreated culture. mRNA levels of CD9 and CD81 were examined by real-time PCR. Data shown are from one representative of three similar experiments. (E) Human monocytic THP-1 cells were treated for 4 h with 1 μg/ml phorbol 12-myristate 13-acetate, allowed to attach to a plate, and then cultured in the absence or presence of increasing concentrations of simvastatin. Levels of CD9, CD63, and CD81 were examined by immunoblotting. (F) Mouse 3T3 fibroblasts were cultured in the absence or presence of increasing concentrations of simvastatin. Levels of CD9, CD63, and CD81 were examined by immunoblotting.
From: Jin Y, Tachibana I, Takeda Y, He P, Kang S, et al. (2013)
Statins Decrease Lung Inflammation in Mice by Upregulating Tetraspanin CD9 in Macrophages.
PLoS ONE 8(9): e73706.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Hamster anti mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in BMDM lysates by western blotting.
Image caption:
The anti-inflammatory effects of statins are CD9-dependent. (A) BMDMs from WT mice were cultured for 24 h in the absence (-) or presence of 3 μM fluvastatin (Fluv) (+), and unstimulated (-) or stimulated for 24 h with 1 μg/ml LPS (+). The cells were lysed, and levels of CD9 and CD81 were examined by immunoblotting. Anti-actin blots show that comparable amounts of protein were loaded in each lane. (B) BMDMs from WT and CD9 KO mice were cultured in the absence or presence of the indicated concentrations of fluvastatin, and stimulated for 18 h with 10 μg/ml LPS (+). Activities of MMP-9 in culture supernatants were analyzed by gelatin zymography. (C) BMDMs from WT and CD9 KO mice were cultured in the absence (vehicle) or presence of 10 μM fluvastatin or simvastatin (Simv), and unstimulated (-) or stimulated for 18 h with 1 μg/ml LPS (+). Concentrations of TNF-α in culture supernatants were measured by ELISA. Each bar represents the mean ± SEM. ⋆P < 0.05; ⋆ ⋆ P < 0.01.
From: Jin Y, Tachibana I, Takeda Y, He P, Kang S, et al. (2013)
Statins Decrease Lung Inflammation in Mice by Upregulating Tetraspanin CD9 in Macrophages. PLoS ONE 8(9): e73706.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Hamster anti mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in RAW264.7 lysates by western blotting.
Image caption:
Blockade of the mevalonate pathway increases CD9 and CD81. (A) RAW264.7 cells were untreated (-) or treated for 48 h with 50 ng/ml TSA (+) in the absence (-) or presence of 50 μM theophylline or 0.5 μM fluvastatin (Fluv) (+). The cells were lysed, and levels of CD9 and CD81 were examined by immunoblotting. Anti-actin blots show that comparable amounts of protein were loaded in each lane. (B) The mevalonate pathway and inhibitors. n-BP, nitrogenous bisphosphonate. (C) RAW264.7 cells were cultured for 24 h in the presence of indicated concentrations of fluvastatin, simvastatin (Simv), zoledronate (Zol), or risedronate (Ris). Levels of CD9 and CD81 were examined by immunoblotting. (D) RAW264.7 cells were cultured for 24 h in the absence (V, vehicle alone) or presence of mevalonate (Mev), farnesyl pyrophosphate (FPP), squalene (Squ), or geranylgeranyl pyrophosphate (GGPP). Although the actin level in the GGPP lane appears to be lower, an equal amount of protein was loaded. (E) RAW264.7 cells were cultured for 24 h in the absence (V) or presence of fluvastatin, zoledronate, farnesyl transferase inhibitor (FTI), or geranylgeranyl transferase inhibitor (GGTI). (F) RAW264.7 cells were untreated (-) or treated with fluvastatin (+) in the absence (V) or presence of mevalonate, FPP, squalene, or GGPP. (G) RAW264.7 cells were untreated (-) or treated with zoledronate (+) in the absence (V) or presence of mevalonate, FPP, squalene, or GGPP. (H) RAW264.7 cells were untreated (-) or treated with fluvastatin (+) in the absence (V) or presence of mevalonate, FPP, squalene, or GGPP and stimulated for 15 min with 0.1 µg/ml LPS (+). The cells were lysed, and levels of IκBα were examined by immunoblotting. (I) RAW264.7 cells were cultured for 24 h in the indicated concentrations of HA1077. Levels of CD9 and CD81 were examined by immunoblotting.
From: Jin Y, Tachibana I, Takeda Y, He P, Kang S, et al. (2013)
Statins Decrease Lung Inflammation in Mice by Upregulating Tetraspanin CD9 in Macrophages.
PLoS ONE 8(9): e73706.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Hamster anti mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in EV lysates by western blotting.
Image caption:
Density fractionation of EVs. The figure shows sucrose gradients of EVs preparations from MLP29 (A) and RH (B). Aliquots of these fractions were used for RNA extraction and protein extraction; the most abundant transcripts were found in the fractions containing typical exosomal markers (Tsg101 or Aip1). RH preparations showed more diversity, with vesicle populations fractionating at different densities.
From: Royo F, Schlangen K, Palomo L, Gonzalez E, Conde-Vancells J, et al. (2013)
Transcriptome of Extracellular Vesicles Released by Hepatocytes.
PLoS ONE 8(7): e68693.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
Hamster anti Mouse CD81 antibody, clone Eat2 (MCA1846) used for the evaluation of CD81 expression in mouse lung lysates by western blotting.
Image caption:
Double deletion of CD9/CD81 results in reduced SIRT1 expression. (a) Immunoblot of the lungs for major aging molecules. Notably, expression of SIRT1, but not that of SIRT6, Klotho, WRN, and ATR, was reduced in DKO lungs. (b) Knockdown of CD9 and CD81 expression with siRNA in epithelial cells additively downregulated the expression of SIRT1. (c) SIRT1 expression by ELISA (n = 4). (d) SIRT1 expression by immunocytochemistry. Bars represent means ± SD; *P <0.05 versus si-cont. Scale bar, 50 μm for (d).
From: Jin Y, et al.
A. Double deletion of tetraspanins CD9 and CD81 in mice leads to a syndrome resembling accelerated aging.
Sci Rep. 2018 Mar 23;8(1):5145.
This is from an open access article distributed under the terms of the Creative Commons Attribution License.
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F WB IP ResetHamster anti Mouse CD81
- Product Type
- Monoclonal Antibody
- Clone
- Eat2
- Isotype
- IgG1
- Specificity
- CD81
Quick Links:
Antibody Quality and 
Unrivaled 
| Hamster anti Mouse CD81 antibody, clone Eat2 recognizes mouse and rat CD81, also known as TAPA-1 or Target of the antiproliferative antibody 1. CD81 is a 236 amino acid ~26 kDa multipass transmembrane protein belonging to the TM4SF family (UniProt: P35762). In rodents CD81 is expressed at much higher levels on resting B cells than on T cells, although increased expression on T cells is found following activation. Hamster anti Mouse CD81 antibody, clone Eat2 induces homotypic aggregation of B cells and inhibits anti Ig and IL-4 induced proliferation (Maecker et al. 2000). Eat 2 requires the presence of both extracellular loops of TAPA-1 for binding. Mice lacking CD81 demonstrate reduced fertility through impaired oocyte-sperm fusion, double knockout CD81-/- CD9-/- mice are completely infertile suggesting complimentary roles in oocyte-sperm fusion (Rubenstein et al. 2006). |
- Target Species
- Mouse
- Species Cross-Reactivity
-
Target Species Cross Reactivity Rat - N.B. Antibody reactivity and working conditions may vary between species.
- Product Form
- Purified IgG - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
0.09% Sodium Azide - Carrier Free
- Yes
- Immunogen
- 38C13, murine B cell line.
- Approx. Protein Concentrations
- Pack Size: 0.1 mgIgG concentration 1 mg/mlPack Size: 25 µg, 0.25 mgIgG concentration 1.0 mg/ml
- Fusion Partners
- Spleen cells from immunised Armenian hamsters were fused with cells of the mouse PX3-Ag.8.653 myeloma cell line.
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
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| Flow Cytometry | |
1/10 Pack Size: 0.1 mg 1/50 Pack Size: 25 µg, 0.25 mg |
1/50 Pack Size: 0.1 mg 1/100 Pack Size: 25 µg, 0.25 mg |
| Immunohistology - Frozen 1 | |
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| Immunohistology - Paraffin | |||
| Immunoprecipitation | |
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| Western Blotting 2 | |
- 1The epitope recognised by this antibody is reported to be sensitive to formaldehyde fixation and tissue processing. Bio-Rad recommends the use of acetone fixation for frozen sections.
- 2Clone Eat2 recognizes mouse CD81 under non-reducing conditions.
Where this antibody has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the antibody for use in their own system using appropriate negative/positive controls.
- Flow Cytometry
- Use 10ul of the suggested working dilution to label 106 cells in 100ul.
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References for CD81 antibody
-
Clark, K.L. et al. (2001) PGRL is a major CD81-associated protein on lymphocytes and distinguishes a new family of cell surface proteins.
J Immunol. 167 (9): 5115-21. -
Maecker, H.T. et al. (2000) Differential expression of murine CD81 highlighted by new anti-mouse CD81 monoclonal antibodies.
Hybridoma 19: 15-22. -
Conde-Vancells, J. et al. (2010) Candidate biomarkers in exosome-like vesicles purified from rat and mouse urine samples.
Proteomics Clin Appl. 4 (4): 416-25. -
Conde-Vancells, J. et al. (2008) Characterization and comprehensive proteome profiling of exosomes secreted by hepatocytes.
J Proteome Res. 7: 5157-66. -
Takeda, Y. et al. (2008) Double deficiency of tetraspanins CD9 and CD81 alters cell motility and protease production of macrophages and causes chronic obstructive pulmonary disease-like phenotype in mice.
J Biol Chem. 283: 26089-97. -
Suzuki, M. et al. (2009) Tetraspanin CD9 negatively regulates lipopolysaccharide-induced macrophage activation and lung inflammation.
J Immunol. 182: 6485-93. -
Ha, C.T. et al. (2005) Binding of pregnancy-specific glycoprotein 17 to CD9 on macrophages induces secretion of IL-10, IL-6, PGE2, and TGF-beta1.
J Leukoc Biol. 77: 948-57. -
Pan, Q. et al. (2011) Hepatic cell-to-cell transmission of small silencing RNA can extend the therapeutic reach of RNA interference (RNAi).
Gut. 61: 1330-9.
View The Latest Product References
-
Jin, Y. et al. (2013) Statins decrease lung inflammation in mice by upregulating tetraspanin CD9 in macrophages.
PLoS One. 8: e73706. -
Royo, F. et al. (2013) Transcriptome of extracellular vesicles released by hepatocytes.
PLoS One. 8: e68693. -
Owens, D.M. and Watt, F.M. (2001) Influence of beta1 integrins on epidermal squamous cell carcinoma formation in a transgenic mouse model: alpha3beta1, but not alpha2beta1, suppresses malignant conversion.
Cancer Res. 61: 5248-54. -
Jin, Y. et al. (2018) Double deletion of tetraspanins CD9 and CD81 in mice leads to a syndrome resembling accelerated aging.
Sci Rep. 8 (1): 5145. -
Sosa, L.J. et al. (2013) Amyloid precursor protein is an autonomous growth cone adhesion molecule engaged in contact guidance.
PLoS One. 8 (5): e64521.
- Synonyms
- TAPA-1
- RRID
- AB_323200
- UniProt
- P35762
- Entrez Gene
- Cd81
- GO Terms
- GO:0000187 activation of MAPK activity
- GO:0005515 protein binding
- GO:0008104 protein localization
- GO:0005887 integral to plasma membrane
- GO:0006661 phosphatidylinositol biosynthetic process
- GO:0030307 positive regulation of cell growth
- GO:0030890 positive regulation of B cell proliferation
- GO:0043128 positive regulation of 1-phosphatidylinositol 4-kinase activity
- GO:0050731 positive regulation of peptidyl-tyrosine phosphorylation
- View More GO Terms
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