Revision 8

#14997Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, IP, IF-F, IF-IC

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

30

Source/Isotype:

Rabbit IgG

UniProt ID:

#Q8NE86

Entrez-Gene Id:

90550

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunofluorescence (Frozen) 1:800 - 1:1600
Immunofluorescence (Immunocytochemistry) 1:1600 - 1:3200

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier free (BSA and azide free) version of this product see product #50261.

Specificity / Sensitivity

MCU (D2Z3B) recognizes endogenous levels of total MCU protein. The antibody non-specifically labels the nucleus in fixed frozen mouse brain by immunofluorescence.

Species Reactivity:

Human, Mouse, Rat, Monkey

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human MCU protein.

Background

The mitochondrial calcium uniporter (MCU) is an inner membrane transport protein that is essential for the regulation of calcium uptake and the maintenance of mitochondrial calcium homeostasis (1). MCU is responsible for the rapid transport of calcium ions across the inner membrane and into the mitochondrial matrix, taking advantage of the membrane potential generated by the electron transport chain. Subsequent release of calcium from the matrix through antiporter proteins regulates a number of biological processes, including signal transduction, bioenergetics, and cell death and survival (2,3). The MCU protein contains a pair of transmembrane domains with both carboxy- and amino-terminal ends within the mitochondrial matrix (3). The surrounding uniporter complex contains a number of proteins that regulate calcium ion transport, including the mitochondrial calcium uniporter regulator 1 (MCUR1), mitochondrial calcium uptake proteins 1 and 2 (MICU1, MICU2), and the essential MCU regulator EMRE (3). MICU1 stabilizes the closed state of the transport complex and preserves the normal mitochondrial calcium concentration below the equilibrium level during resting conditions (4). The MCUR1 protein is an essential regulator of calcium uptake, with decreased ion transport in cells with reduced MCUR1 expression (2). EMRE is also essential for MCU transport function where it functions as bridge between MCU uniporter activity and the calcium-sensing MICU1/2 proteins (5).

  1. Patron, M. et al. (2013) J Biol Chem 288, 10750-8.
  2. Mallilankaraman, K. et al. (2012) Nat Cell Biol 14, 1336-43.
  3. Marchi, S. and Pinton, P. (2014) J Physiol 592, 829-39.
  4. Mallilankaraman, K. et al. (2012) Cell 151, 630-44.
  5. Sancak, Y. et al. (2013) Science 342, 1379-82.

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

WB: Western Blotting IP: Immunoprecipitation IF-F: Immunofluorescence (Frozen) 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 All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
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 8
#14997

MCU (D2Z3B) Rabbit mAb

Western Blotting Image 1: MCU (D2Z3B) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using MCU (D2Z3B) Rabbit mAb.
Immunoprecipitation Image 1: MCU (D2Z3B) Rabbit mAb Expand Image
Immunoprecipitation of MCU from C6 cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or MCU (D2Z3B) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using MCU (D2Z3B) Rabbit mAb.
Immunofluorescence Image 1: MCU (D2Z3B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse retina, labeled with MCU (D2Z3B) Rabbit mAb #14997 (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182 (right, red) and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 2: MCU (D2Z3B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of fixed frozen mouse colon, labeled with MCU (D2Z3B) Rabbit mAb #14997 (left, green). Free secondary binding sites were then blocked with Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 prior to co-labeling with Ras (E4K9L) Rabbit mAb (Alexa Fluor® 647 Conjugate) #37182 (right, red) and ProLong® Gold Antifade Reagent with DAPI #8961 (right, blue).
Immunofluorescence Image 1: MCU (D2Z3B) Rabbit mAb Expand Image
Confocal immunofluorescent analysis of HCT 116 cells, labeled with MCU (D2Z3B) Rabbit mAb #14997 (left, green) and co-labeled with DyLight 650 Phalloidin #12956 (right, red), and DAPI #4083 (right, blue).