Revision 1

#14256Store at -20C

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

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

WB

REACTIVITY:

H M R Mk

SENSITIVITY:

Endogenous

MW (kDa):

14

Source/Isotype:

Rabbit IgG

UniProt ID:

#P60520

Entrez-Gene Id:

11345

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000

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.

Specificity / Sensitivity

GABARAPL2 (D1W9T) Rabbit mAb recognizes endogenous levels of total GABARAPL2 protein. Bands of unknown origin are detected at 80 and 110 kDa in some cell lines. This antibody has a preference for the Type I form of GABARAPL2.

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 GABARAPL2 protein.

Background

GABAA receptor associated protein (GABARAP) is an Atg8 family protein with a key role in autophagy, which was originally discovered as a protein associated with the GABAA receptor regulating receptor trafficking to the plasma membrane (1). Proteins in this family, including microtubule-associated protein light chain 3 (LC3) and GATE-16 (GABARAPL2), become incorporated into the autophagosomal membranes following autophagic stimuli such as starvation (2). Like the other family members, GABARAP is cleaved at its carboxyl terminus, which leads to conjugation by either of the phospholipids phosphatidylethanolamine or phosphatidylserine (3,4). This processing converts GABARAP from a type I to a type II membrane bound form involved in autophagosome biogenesis. Processing of GABARAP involves cleavage by Atg4 family members (5,6) followed by conjugation by the E1 and E2 like enzymes Atg7 and Atg3 (7,8). GABARAPL1/GEC1, a protein that is highly related to GABARAP, was identified as an estrogen inducible gene, and is also associated with autophagosomes (9-11).
GABARAPL2/GATE-16 was identified as a modulator of membrane transport, interacting with N-ethylmaleimide senstive factor (NSF) and the Golgi v-SNARE GOS-28 (12). In addition, GABARAPL2 interacts with OSBP-related protein 7 (ORP7), the GTPase GIMAP6, and the calcium channel TRPML3. (13-15)

  1. Wang, H. et al. (1999) Nature 397, 69-72.
  2. Shpilka, T. et al. (2011) Genome Biol 12, 226.
  3. Kabeya, Y. et al. (2004) J Cell Sci 117, 2805-12.
  4. Sou, Y.S. et al. (2006) J Biol Chem 281, 3017-24.
  5. Tanida, I. et al. (2004) J Biol Chem 279, 36268-76.
  6. Hemelaar, J. et al. (2003) J Biol Chem 278, 51841-50.
  7. Tanida, I. et al. (2001) J Biol Chem 276, 1701-6.
  8. Tanida, I. et al. (2002) J Biol Chem 277, 13739-44.
  9. Chakrama, F.Z. et al. (2010) Autophagy 6, 495-505.
  10. Pellerin, I. et al. (1993) Mol Cell Endocrinol 90, R17-21.
  11. Vernier-Magnin, S. et al. (2001) Biochem Biophys Res Commun 284, 118-25.
  12. Sagiv, Y. et al. (2000) EMBO J 19, 1494-504.
  13. Zhong, W. et al. (2011) Exp Cell Res 317, 2353-63.
  14. Pascall, J.C. et al. (2013) PLoS One 8, e77782.
  15. Choi, S. and Kim, H.J. (2014) Biochem Biophys Res Commun 443, 56-61.

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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting

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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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
#14256

GABARAPL2 (D1W9T) Rabbit mAb

Western Blotting Image 1: GABARAPL2 (D1W9T) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® GABARAPL2 siRNA I # 14239 (+) or SignalSilence® II GABARAPL2 siRNA II #14246 (+), using GABARAPL2 (D1W9T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The GABARAPL2 (D1W9T) Rabbit mAb confirms silencing of GABARAPL2, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.
Western Blotting Image 2: GABARAPL2 (D1W9T) Rabbit mAb Expand Image
Western blot analysis of extracts from various cell lines using GABARAPL2 (D1W9T) Rabbit mAb.
Western Blotting Image 3: GABARAPL2 (D1W9T) Rabbit mAb Expand Image
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human GABARAP (hGABARAP-Myc/DDK; +), human GABARAPL1 (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 (hGABARAPL2-Myc/DDK; +), using GABARAPL2 (D1W9T) Rabbit mAb.
Western Blotting Image 4: GABARAPL2 (D1W9T) Rabbit mAb Expand Image
Western blot analysis of extracts from A172 and HeLa cells, untreated (-) or treated overnight with chloroquine (50 μM) (+), using GABARAPL2 (D1W9T) Rabbit mAb (upper) or β-Actin (D6A8) #8457 (lower).