#8648 Parkinson's Research Antibody Sampler Kit
|DJ-1 (D29E5) XP® Rabbit mAb #5933||20 µl||WB, IP, IF-IC||H, M, R, Hm, Mk||22||Rabbit IgG|
|LRRK2 (D18E12) Rabbit mAb #13046||20 µl||WB, IP||H, M, R||290||Rabbit IgG|
|Parkin (Prk8) Mouse mAb #4211||20 µl||WB, IP||H, M, R||50||Mouse IgG2b|
|PINK1 (D8G3) Rabbit mAb #6946||20 µl||WB, IP||H||60, 50||Rabbit IgG|
|α-Synuclein (D37A6) XP® Rabbit mAb #4179||20 µl||WB, IP, IHC-P, IF-F||M, R||18||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||WB||Goat|
|Anti-mouse IgG, HRP-linked Antibody #7076||100 µl||WB||Horse|
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), IHC-P=Immunohistochemistry (Paraffin), IF-F=Immunofluorescence (Frozen)
Reactivity Key: H=Human, M=Mouse, R=Rat, Hm=Hamster, Mk=Monkey
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.
Western blot analysis of extracts from PC12 cells, fetal rat brain and mouse brain, using Parkin (Prk8) Mouse mAb.
Western blot analysis of extracts from mouse and rat brain using α-Synuclein (D37A6) XP® Rabbit mAb.
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).
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.
Western blot analysis of extracts from U-87 MG and A172 cells, and mouse brain using LRRK2 (D18E12) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse brain using α-Synuclein (D37A6) XP® Rabbit mAb.
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).
Western blot analysis of extracts from HeLa cells, untreated (-) or treated with CCCP (10 μM, 24 hr; +), using PINK1 (D8G3) Rabbit mAb.
Confocal immunofluorescent analysis of normal rat cerebellum, hippocampus and striatum using α-Synuclein (D37A6) XP® Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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).
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DRAQ5 is a registered trademark of Biostatus Limited.
XP is a registered trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.