Revision 2

#8648Store at -20C

1 Kit

(5 x 20 microliters)

Cell Signaling Technology

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

Support: 877-678-TECH (8324)

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3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
DJ-1 (D29E5) XP® Rabbit mAb 5933 20 µl 22 kDa Rabbit IgG
LRRK2 (D18E12) Rabbit mAb 13046 20 µl 290 kDa Rabbit IgG
Parkin (Prk8) Mouse mAb 4211 20 µl 50 kDa Mouse IgG2b
PINK1 (D8G3) Rabbit mAb 6946 20 µl 60, 50 kDa Rabbit IgG
α-Synuclein (D37A6) Rabbit mAb 4179 20 µl 18 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

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

Description

The Parkinson's Research Antibody Sampler Kit provides an economical means of detecting target proteins related to Parkinson's disease. The kit contains enough primary and secondary antibody to perform two western blots per primary.

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

Parkinson’s disease (PD), the second most common neurodegenerative disease after Alzheimer’s, is a progressive movement disorder characterized by rigidity, tremors, and postural instability. The pathological hallmark of PD is progressive loss of dopaminergic neurons in the substantia nigra of the ventral midbrain and the presence of intracellular Lewy bodies in surviving neurons of the brain stem (1). Research studies have shown that various genes and loci (α-synuclein/PARK1 and 4, parkin/PARK2, UCH-L1/PARK5, PINK1/PARK6, DJ-1/PARK7, LRRK2/PARK8, synphilin-1, and NR4A2) are genetically linked to PD (2).

α-Synuclein, a 140 amino acid protein expressed abundantly in the brain, is a major component of aggregates found in Lewy bodies (3). Parkin is involved in protein degradation through the ubiquitin-proteasome pathway, and investigators have shown that mutations in Parkin cause early onset of PD (4). In the case of autosomal recessive juvenile Parkinsonism (AR-JP), deletions have been found on chromosome 6 in the Parkin gene (5). PTEN induced putative kinase 1 (PINK1) is a mitochondrial serine/threonine kinase involved in the normal function and integrity of mitochondria, as well as a reduction of cytochrome c release from mitochondria (6-8). PINK1 phosphorylates Parkin and promotes its translocation to mitochondria (7). Mutations of PINK1 are associated with loss of protective function, mitrochondrial dysfunction, aggregation of α-synuclein, and proteasome dysfunction (6,8). DJ-1 is involved in multiple cellular functions; it has been shown to cooperate with Ras to increase cell transformation, to regulate transcription of the androgen receptor, and may function as an indicator of oxidative stress, while loss-of-function mutations in DJ-1 cause early onset of PD (9-12). Dopamine D2 receptor-mediated functions are greatly impaired in DJ-1 (-/-) mice, resulting in reduced long-term depression (13). Leucine-rich repeat kinase 2 (LRRK2) contains amino-terminal leucine-rich repeats (LRR), a Ras-like small GTP binding protein-like (ROC) domain, an MLK protein kinase domain, and a carboxy-terminal WD40-repeat. At least 20 LRRK2 mutations have been linked to PD (14). The most prevalent mutation, G2019S, causes increased LRRK2 kinase activity, leading to progressive neurite loss and decreased neuronal survival (15).

  1. Fahn, S. (2003) Ann N Y Acad Sci 991, 1-14.
  2. Moore, D.J. et al. (2005) Annu Rev Neurosci 28, 57-87.
  3. Goldberg, M.S. and Lansbury, P.T. (2000) Nat Cell Biol 2, E115-9.
  4. Borrelli, E. (2005) Neuron 45, 479-81.
  5. Polymeropoulos, M.H. et al. (1997) Science 276, 2045-7.
  6. Liu, W. et al. (2009) PLoS One 4, e4597.
  7. Kim, Y. et al. (2008) Biochem Biophys Res Commun 377, 975-80.
  8. Petit, A. et al. (2005) J Biol Chem 280, 34025-32.
  9. Bonifati, V. et al. (2003) Science 299, 256-9.
  10. Nagakubo, D. et al. (1997) Biochem Biophys Res Commun 231, 509-13.
  11. Takahashi, K. et al. (2001) J Biol Chem 276, 37556-63.
  12. Mitsumoto, A. and Nakagawa, Y. (2001) Free Radic Res 35, 885-93.
  13. Goldberg, M.S. et al. (2005) Neuron 45, 489-96.
  14. Mata, I.F. et al. (2006) Trends Neurosci 29, 286-93.
  15. MacLeod, D. et al. (2006) Neuron 52, 587-93.

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

    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 2
    #8648

    Parkinson's Research Antibody Sampler Kit

    Parkinson's Research Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of lysates (0.1 mg/mL) from C6 cells using DJ-1 (D29E5) XP Rabbit mAb #5933. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    Parkinson's Research Antibody Sampler Kit: Image 2 Expand Image
    Western blot analysis of extracts from U-87 MG and A172 cells, and mouse brain using LRRK2 (D18E12) Rabbit mAb. 
    Parkinson's Research Antibody Sampler Kit: Image 3 Expand Image
    Western blot analysis of extracts from mouse and rat brain using α-Synuclein (D37A6) Rabbit mAb.
    Parkinson's Research Antibody Sampler Kit: Image 4 Expand Image
    Western blot analysis of extracts from PC12 cells, fetal rat brain and mouse brain, using Parkin (Prk8) Mouse mAb.
    Parkinson's Research Antibody Sampler Kit: Image 5 Expand Image
    Western blot analysis of extracts from MEF wild-type, MEF DJ-1 (-/-), HeLa, and C6 cells using DJ-1 (D29E5) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). (MEF wild-type and MEF DJ-1 (-/-) cells were kindly provided by Dr. Philipp Kahle, University of Tübingen, Germany).
    Parkinson's Research Antibody Sampler Kit: Image 6 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a cDNA construct expressing full-length human PINK1 (hPINK1, +) using PINK1 (D8G3) Rabbit mAb.
    Parkinson's Research Antibody Sampler Kit: Image 7 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.
    Parkinson's Research Antibody Sampler Kit: Image 8 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.
    Parkinson's Research Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of A549 extracts from WT (left) or LRRK2 KO (right) using LRRK2 (D18E12) Rabbit mAb. Membranes stained with Ponceau S for total protein normalization (lower). These data were provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibodies, as a companion to validation data generated by CST scientists.
    Parkinson's Research Antibody Sampler Kit: Image 10 Expand Image
    Immunoprecipitation of Parkin protein from PC12 extracts. Lane 1 is 10% input, lane 2 is Mouse (E7Q5L) mAb IgG2b Isotype Control #53484, and lane 3 is Parkin (Prk8) Mouse mAb. Western blot analysis was performed using Parkin Antibody #2132. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as a secondary antibody.
    Parkinson's Research Antibody Sampler Kit: Image 11 Expand Image
    Confocal immunofluorescent analysis of MEF wild-type (left) or MEF DJ-1 (-/-) (right) cells using DJ-1 (D29E5) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). (MEF wild-type and MEF DJ-1 (-/-) cells were kindly provided by Dr. Philipp Kahle, University of Tübingen, Germany).
    Parkinson's Research Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from HeLa cells, untreated (-) or treated with CCCP (10 μM, 24 hr; +), using PINK1 (D8G3) Rabbit mAb.
    Parkinson's Research Antibody Sampler Kit: Image 13 Expand Image
    Immunoprecipitation of LRRK2 from mouse brain extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is LRRK2 (D18E12) Rabbit mAb. Western blot analysis was performed using LRRK2 (D18E12) Rabbit mAb. Anti-rabbit, HRP-linked Antibody #7074 was used as a secondary antibody.
    Parkinson's Research Antibody Sampler Kit: Image 14 Expand Image
    Confocal immunofluorescent analysis of normal rat cerebellum, hippocampus and striatum using α-Synuclein (D37A6) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).