Revision 1

#48046Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

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

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
SQSTM1/p62 (D5E2) Rabbit mAb 8025 20 µl 62 kDa Rabbit IgG
Optineurin (D2L8S) Rabbit mAb 58981 20 µl 75 kDa Rabbit IgG
NBR1 (D2E6) Rabbit mAb 9891 20 µl 120 kDa Rabbit IgG
NDP52 (D1E4A) Rabbit mAb 60732 20 µl 52, 60 kDa Rabbit IgG
TAX1BP1 (D1D5) Rabbit mAb 5105 20 µl 92 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The SQSTM1/p62-like Receptor Antibody Sampler Kit provides an economical means of detecting members of the SQSTM1/p62-like Receptor (SLR) family. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

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.

Background

Autophagy is a catabolic process for the autophagosome-lysosomal degradation of bulk cytoplasmic contents (1,2). Selective autophagy targets the degradation of distinct sets of substrates and organelles and can occur through the utilization of a number of autophagy cargo receptors (3-5). Autophagy cargo receptors contain an LC3-interacting region (LIR) required for interaction with Atg8/LC3 family members targeted to the autophagosome. SQSTM1/p62-like receptors (SLRs) are a family of autophagy cargo receptors that contain domains for binding to ubiquitin. This family includes prototypical member SQSTM1/p62, NBR1, NDP52, Optineurin, and TAX1BP1. Targets of SLRs include ubiquitylated protein aggregates (aggrephagy), organelles such as mitochondria (mitoophagy) and peroxisomes (pexophagy), and intracellular bacteria (xenophagy).

Upon binding of cargo to these receptors, the complex is delivered to the autophagosome where both the cargo and receptor are degraded through the autophagic process. While some redundancy may exist among SLR family members, they can have unique activities. Many SLRs can have additional roles as scaffolding proteins for various signaling pathways. For example, SQSTM1/p62 interacts with KEAP1, a cytoplasmic inhibitor of NRF2, a key transcription factor involved in cellular responses to oxidative stress (6). Thus, accumulation of SQSTM1/p62 can lead to an increase in NRF2 activity.

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  3. Birgisdottir, Å.B. et al. (2013) J Cell Sci 126, 3237-47.
  4. Xu, Z. et al. (2015) Acta Biochim Biophys Sin (Shanghai) 47, 571-80.
  5. Mancias, J.D. and Kimmelman, A.C. (2016) J Mol Biol 428, 1659-80.
  6. Komatsu, M. et al. (2010) Nat Cell Biol 12, 213-23.

Background References

    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
    #48046

    SQSTM1/p62-like Receptor Antibody Sampler Kit

    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 1 Expand Image
    Western blot analysis of extracts from various cell lines using TAX1BP1 (D1D5) Rabbit mAb.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from various cell lines using Optineurin (D2L8S) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from various cell lines using NDP52 (D1E4A) Rabbit mAb.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 4 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from HeLa cells (lane 1) or SQSTM1 knock-out cells (lane 2) using SQSTM1/p62 (D5E2) Rabbit mAb #8025 (upper), and β-actin (D6A8) Rabbit mAb #8457 (lower). The absence of signal in the SQSTM1 knock-out HeLa cells confirms specificity of the antibody for SQSTM1.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from various cell lines using NBR1 (D2E6) Rabbit mAb.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of extracts from 293T cells, either mock transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding the full-length isoform 1 of human TAX1BP1 (hTAX1BP1-Myc/DDK, +), using TAX1BP1 (D1D5) Rabbit mAb.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® SQSTM1/p62 siRNA I #6394 (+) or SignalSilence® SQSTM1/p62 siRNA II #6399 (+), using SQSTM1/p62 (D5E2) Rabbit mAb (upper) or α-Tubulin (11H10) Rabbit mAb #2125 (lower). The SQSTM1/p62 (D5E2) Rabbit mAb confirms silencing of SQSTM1/p62 expression, while the α-Tubulin (11H10) Rabbit mAb is used as a loading control.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 9 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with bafilomycin A (0.2 μM, 18 hr; right), using NBR1 (D2E6) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from various cell lines using SQSTM1/p62 (D5E2) Rabbit mAb.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 11 Expand Image
    Flow cytometric analysis of MCF7 cells using NBR1 (D2E6) Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from SK-MEL-2 cells, untreated (-) or starved overnight in Earle's Balanced Salt Solution (EBSS) (+), using SQSTM1/p62 (D5E2) Rabbit mAb.
    SQSTM1/p62-like Receptor Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a tagged human SQSTM1/p62 construct (+), using SQSTM1/p62 (D5E2) Rabbit mAb.