Revision 3
#98327
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IF-IC, FC-FP, FC-L
Reactivity:
M R
Sensitivity:
Endogenous
MW (kDa):
22-27
Source/Isotype:
Rabbit IgG
UniProt ID:
#P40240
Entrez-Gene Id:
12527
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunofluorescence (Immunocytochemistry) | 1:50 |
Flow Cytometry (Fixed/Permeabilized) | 1:50 - 1:200 |
Flow Cytometry (Live) | 1:50 - 1:200 |
Storage
Specificity/Sensitivity
CD9 (E8L5J) Rabbit mAb recognizes endogenous levels of total CD9 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu119 within the extracellular domain of mouse CD9 protein.
Background
The CD9 antigen belongs to the tetraspanin family of cell surface glycoproteins, and is characterized by four transmembrane domains, one short extracellular domain (ECL1), and one long extracellular domain (ECL2). Tetraspanins interact with a variety of cell surface proteins and intracellular signaling molecules in specialized tetraspanin-enriched microdomains (TEMs), where they mediate a range of processes including adhesion, motility, membrane organization, and signal transduction (1). Research studies demonstrate that CD9 expression on the egg is required for gamete fusion during fertilization (2-4). CD9 was also shown to play a role in dendritic cell migration, megakaryocyte differentiation, and homing of cord blood CD34+ hematopoietic progenitors to the bone marrow (5-7). In addition, downregulation of CD9 expression is associated with poor prognosis and progression of several types of cancer (8-10). Additional research identified CD9 as an abundant component of exosomes, and may play some role in the fusion of these secreted membrane vesicles with recipient cells (11).
Background References
- Hemler, M.E. (2005) Nat Rev Mol Cell Biol 6, 801-11.
- Le Naour, F. et al. (2000) Science 287, 319-21.
- Miyado, K. et al. (2000) Science 287, 321-4.
- Kaji, K. et al. (2000) Nat Genet 24, 279-82.
- Mantegazza, A.R. et al. (2004) Blood 104, 1183-90.
- Clay, D. et al. (2001) Blood 97, 1982-9.
- Leung, K.T. et al. (2011) Blood 117, 1840-50.
- Miyake, M. et al. (1995) Cancer Res 55, 4127-31.
- Higashiyama, M. et al. (1995) Cancer Res 55, 6040-4.
- Uchida, S. et al. (1999) Br J Cancer 79, 1168-73.
- Théry, C. et al. (1999) J Cell Biol 147, 599-610.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.
Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.
Revision 3
#98327
CD9 (E8L5J) Rabbit mAb
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length mouse CD9 protein (mCD9-Myc/DDK; +), using CD9 (E8L5J) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
Western blot analysis of extracts from various cell lines using CD9 (E8L5J) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Absence of signal in BA/F3 cells is predicted from RNAseq data and confirms the specificity of the antibody.
Confocal immunofluorescent analysis of RAW 264.7 cells (left, positive) or BA/F3 cells (right, negative) using CD9 (E8L5J) Rabbit mAb (green) and DAPI #4083 (blue).
Revision 3
#98327
CD9 (E8L5J) Rabbit mAb
Flow cytometric analysis of live BA/F3 cells (blue, negative) and JAWSII cells (green, positive) using CD9 (E8L5J) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow cytometric analysis of fixed and permeabilized BA/F3 cells (blue, negative) and JAWSII cells (green, positive) using CD9 (E8L5J) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.