Cell Signaling Technology

Product Pathways - Neuroscience

Parkinson's Research Antibody Sampler Kit #8648

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
DJ-1 (D29E5) XP® Rabbit mAb #5933 40 µl W IP IF-IC H M R Hm Mk 22 Rabbit IgG
LRRK2 Antibody #5559 40 µl W IP H M R 290 Rabbit
Parkin (Prk8) Mouse mAb #4211 40 µl W IP H M R 50 Mouse IgG2b
PINK1 (D8G3) Rabbit mAb #6946 40 µl W IP H 60, 50 Rabbit IgG
α-Synuclein (D37A6) XP® Rabbit mAb #4179 40 µl W IP IHC-P IF-F M R 18 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat
Anti-mouse IgG, HRP-linked Antibody #7076 100 µl Horse

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-F=Immunofluorescence (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

DJ-1 (D29E5) XP® Rabbit mAb, LRRK2 Antibody, Parkin (Prk8) Mouse mAb, and PINK1 (D8G3) Rabbit mAb recognize endogenous levels of respective target proteins. α-Synuclein (D37A6) XP® Rabbit mAb recognizes endogenous levels of the α isoform of synuclein protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse and rat brain using α-Synuclein (D37A6) XP® Rabbit mAb #4179.

Western Blotting

Western Blotting

Western blot analysis of extracts from PC12 cells, fetal rat brain and mouse brain using Parkin (Prk8) Mouse mAb #4211.

Western Blotting

Western Blotting

Western blot analysis of extracts from mouse LRRK2-/- and wild-type tissue using LRRK2 Antibody #5559 (upper) or TrkB (80E3) Rabbit mAb #4603 (lower). (Mouse wild-type and LRRK2-/- brains were kindly provided by Dr. Jie Shen, Brigham and Women's Hospital and Harvard Medical School, Boston, MA).


Western Blotting

Western Blotting

Western blot analysis of extracts from MEF wild-type, MEF DJ-1 (-/-), HeLa, and C6 cells using DJ-1 (D29E5) XP® Rabbit mAb #5933 (upper) or 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).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells either untreated (-) or treated with CCCP (10 μM, 24 hr; +), using PINK1 (D8G3) Rabbit mAb #6946.

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 four western blots per primary.

Source / Purification

Monoclonal antibodies are produced by immuninzing animals with a recombinant protein specific to the carboxy terminus of Parkin protein, a synthetic peptide corresponding to residues surrounding Pro140 of human PINK1 protein, a synthetic peptide corresponding to residues surrounding Lys148 of human DJ-1 protein, or a synthetic peptide corresponding to residues surrounding Glu105 of mouse α-synuclein protein.Polyclonal antibodies are produced by immuninzing animals with a synthetic peptide corresponding to residues surrounding Leu1848 of human LRRK2 protein. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.

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.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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