Parkinson's Research Antibody Sampler Kit
Catalog# K2003
Parkinson's Research Antibody Sampler Kit
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WB
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Human
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Mouse
General Overview
Kit Components
| Product Includes | Specification | Application | Reactivity | Mw |
|---|---|---|---|---|
| PARK7/DJ1[ET1611-45] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human,Mouse,Rat | Predicted band size: 20 kDa |
| LRRK2[HA722932] | 20µl | IHC-P,IF-Tissue,IHC-Fr | Human,Mouse,Rat | Predicted band size: 286 kDa |
| Parkin[HA722952] | 20µl | WB,IHC-P,IF-Tissue,IF-Cell,FC,IP,IHC-Fr | Human,Mouse,Rat,Monkey | Predicted band size: 52 kDa |
| PINK1[HA723021] | 20µl | WB,IHC-P,IHC-Fr | Human,Mouse,Rat | Predicted band size: 63 kDa |
| Alpha-Synuclein[ET7107-31] | 20µl | WB,IF-Cell,IF-Tissue,IHC-P,IP,FC | Human | Predicted band size: 14 kDa |
| Alpaca anti-Rabbit IgG Fc, Recombinant VHH[HA1031] | 100µl | IP,ELISA,IHC-P,WB | Rabbit |
Product Description
Parkinson's Research Antibody Sampler Kit designed to provide you with a variety of trial-size antibodies in a convenient and cost-effective format. The kit can be used to detect some Parkinson's protein. And also includes secondary reagent for detection of these antibodies.
Product Features
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Background
α-Synuclein, a 140 amino acid protein expressed abundantly in the brain, is a major component of aggregates found in Lewy bodies. Parkin is involved in protein degradation through the ubiquitin-proteasome pathway, and investigators have shown that mutations in Parkin cause early onset of PD. </br>In the case of autosomal recessive juvenile Parkinsonism (AR-JP), deletions have been found on chromosome 6 in the Parkin gene . Mutations of PINK1 are associated with loss of protective function, mitrochondrial dysfunction, aggregation of α-synuclein, and proteasome dysfunction. 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. 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.
Data Links
Background References
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.
Images
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Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: SH-SY5Y cell lysate
Lane 3: HepG2 cell lysate
Lane 4: Mouse brain tissue lysate
Lane 5: Rat brain tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 20 kDa
Observed band size: 20 kDa
Exposure time: 20 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-45) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Western blot analysis of PARK7/DJ1 on different lysates with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/1,000 dilution.
Lane 1: A549-si NT cell lysate
Lane 2: A549-si PARK7/DJ1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 20 kDa
Observed band size: 20 kDa
Exposure time: 30 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1611-45) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunofluorescence analysis of frozen rat brain tissue with Rabbit anti-LRRK2 antibody (HA722932) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722932, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue). -
Immunocytochemistry analysis of HeLa cells labeling PARK7/DJ1 with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/100 dilution.
Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PARK7/DJ1 antibody (ET1611-45) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Western blot analysis of Parkin on different lysates with Rabbit anti-Parkin antibody (HA722952) at 1/2,000 dilution.
Lane 1: SH-SY5Y cell lysate (20 µg/Lane)
Lane 2: 293T cell lysate (20 µg/Lane)
Lane 3: COS-1 cell lysate (20 µg/Lane)
Lane 4: Neuro-2a cell lysate (20 µg/Lane)
Lane 5: C6 cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (30 µg/Lane)
Lane 7: Rat brain tissue lysate (30 µg/Lane)
Predicted band size: 52 kDa
Observed band size: 52 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722952) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Application: IHC-Fr
Species: Mouse
Site: Cerebral cortex
Sample: Frozen section
Antibody concentration: 1/500
Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. -
Application: IHC-Fr
Species: Rat
Site: Cerebral cortex
Sample: Frozen section
Antibody concentration: 1/500
Antigen retrieval: The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven. -
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Parkin antibody (HA722952) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722952) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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