Revision 1
#27748
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP, IF-IC
Reactivity:
H
Sensitivity:
Endogenous
MW (kDa):
35-40, 65
Source/Isotype:
Rabbit IgG
UniProt ID:
#Q9GZU1
Entrez-Gene Id:
57192
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Immunoprecipitation | 1:200 |
| Immunofluorescence (Immunocytochemistry) | 1:200 - 1:800 |
Storage
Specificity/Sensitivity
Mucolipin-1 (F8F9Q) Rabbit mAb recognizes endogenous levels of total mucolipin-1 protein. This antibody detects the carboxyl terminal fragment produced following posttranslational cleavage.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human mucolipin-1 protein.
Background
Mucolipin-1 (MCOLN1, TRPML1), a member of the transient receptor potential ion channel superfamily, is a lysosomal/late endosome Ca2+ efflux channel that is important in lysosomal biogenesis (1,2). Mutations in the mucolipin-1 gene are responsible for the human lysosomal storage disease mucolipidosis type IV (MLIV), an autosomal recessive neurodegenerative disease (3). Mucolipin-1 is synthesized as a 580-amino acid protein with six transmembrane domains but can undergo cleavage in the long luminal loop between the first two transmembrane domains (4,5). Mucolipin-1 activity has been linked to autophagy and lysosomal biogenesis. Calcium influx through mucolipin-1 activates CaMKKβ and AMPK, which activate the autophagy kinase ULK1 (6). Calcium influx is also associated with activation of the calcium-dependent phosphatase calcineurin which can trigger activation of TFEB, a key transcription factor regulating autophagy and lysosomal biogenesis (7,8). Mucolipin-1 activity is upregulated by cellular stress, including reactive oxygen species (ROS) (7). Phosphorylation of mucolipin-1 by mTOR results in reduced channel activity and autophagic flux (9). Loss of mucolipin-1 or pharmacological inhibition suppresses autophagy and can reduce tumor growth and metastasis in non-small cell lung cancer and triple-negative breast cancer, suggesting the use of mucolipin-1 antagonists for cancer therapy (10-12).
Background References
- Zeevi, D.A. et al. (2007) Biochim Biophys Acta 1772, 851-8.
- Cheng, X. et al. (2010) FEBS Lett 584, 2013-21.
- Mirabelli-Badenier, M. et al. (2015) Metab Brain Dis 30, 681-6.
- Kiselyov, K. et al. (2005) J Biol Chem 280, 43218-23.
- Miedel, M.T. et al. (2006) J Biol Chem 281, 12751-9.
- Scotto Rosato, A. et al. (2019) Nat Commun 10, 5630.
- Zhang, X. et al. (2016) Nat Commun 7, 12109.
- Zhang, X. et al. (2016) Autophagy 12, 1954-1955.
- Onyenwoke, R.U. et al. (2015) Biochem J 470, 331-42.
- Yin, C. et al. (2019) Cancer Manag Res 11, 8607-8617.
- Xu, M. et al. (2019) Cell Calcium 79, 80-88.
- Yang, Y. et al. (2020) Cells 9, 2682-2688.
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 IP: Immunoprecipitation IF-IC: Immunofluorescence (Immunocytochemistry)
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.
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Limited Uses
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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 1
Western blot analysis of extracts from various cell lines using Mucolipin-1 (F8F9Q) Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). Low expression of mucolipin-1 protein in MCF7 and Ramos cells is consistent with the predicted expression pattern.
Immunoprecipitation of mucolipin-1 protein from TT cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Mucolipin-1 (F8F9Q) Rabbit mAb. Western blot analysis was performed using Mucolipin-1 (F8F9Q) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as a secondary antibody.
Confocal immunofluorescent analysis of TT cells (left, high-expressing) and MCF7 cells (right, low-expressing) cells using Mucolipin-1 (F8F9Q) Rabbit mAb (green), β-Actin (8H10D10) Mouse mAb #3700 (red), and DAPI #4083 (blue).
