PARK7/DJ1 antibody (AA 1-187)

Catalog No. ABIN5542007

This Mouse Monoclonal antibody specifically detects PARK7/DJ1 in WB. It exhibits reactivity toward Human.

Quick Overview for PARK7/DJ1 antibody (AA 1-187) (ABIN5542007)

Target: PARK7/DJ1 (PARK7) (Parkinson Protein 7 (PARK7))

Reactivity: Human

Host: Mouse

Clonality: Monoclonal

Conjugate: This PARK7/DJ1 antibody is un-conjugated

Application: Western Blotting (WB)

Clone: 3E8

Product Details anti-PARK7/DJ1 Antibody

Binding Specificity: AA 1-187

Specificity: This antibody reacts with DJ-1.

Cross-Reactivity (Details): Does not react with Mouse (NIH/3T3, P19, BaF/3) and Rat (PC12).

Immunogen: Recombinant full-length human DJ-1 (1-187 a.a.)

Isotype: IgG1

Application Details

Application Notes: Western blot: 1-10 μg/mL for chemiluminescence detection system.

Protocol: SDS-PAGE & Western Blotting 1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the Lysis buffer to make an 8 mg/mL solution. 3) Mix the sample with an equal volume of Laemmli's sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μ L of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20 % MeOH). See the manufacture's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature, or overnight at 4 °C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7.2 containing 1 % skimmed milk as suggested in the APPLICATIONS for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 8) Wash the membrane with PBS-T [0.05 % Tween-20 in PBS] (5 minutes x 6 times). 9) Incubate the membrane with the 1:10,000 HRP-conjugated anti-mouse IgG diluted with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS-T (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 1 minute. Develop the film as usual. The conditions for exposure and development may vary. (Positive controls for Western blotting Jurkat, Raji, HeLa)

Restrictions: For Research Use only

Handling

Format: Liquid

Buffer: Buffer System: Protein A agarose, PBS containing 50 % glycerol, pH 7.2. Contains no preservatives.

Preservative: Without preservative

Storage: -20 °C

Storage Comment: Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.

Target Details for PARK7/DJ1

Target: PARK7/DJ1 (PARK7) (Parkinson Protein 7 (PARK7))

Alternative Name: dj-1,park7

Background: DJ-1 (PARK7/CAP1/RS) was originally cloned as a putative oncogene capable of transforming NIH/3T3 cells in cooperation with h-ras, a protein expressed in sperm, and a regulator of RNA-protein interactions. DJ-1 has also been isolated as a gene associated with autosomal ea rly-onset Parkinson's disease (PD). Taken together, DJ-1 appears to be involved in diverse biological processes. First, several lines of evidence suggest that DJ-1 plays a role in the oxidative stress response. In cultured mammalian cells, DJ-1 quenches reactive oxygen species and is converted into a variant with a more acidic isoelectric poin t. Therefore, DJ-1 protects against reactive oxygen specie s-induced cell death, and its suppression with small interfe ring RNA (siRNA) sensitizes cells to such insults. Second, DJ-1 modulates transcription through interaction with DJ-1-binding protein as well as with protein inhibitor of activated STAT (PIAS). The latter modulates the activity of various transcription factors. Third, DJ-1 has been recognized as a regulatory subunit of an RNA-binding protein. Fourth, DJ-1, which is structurally related to the molecular chaperone Hsp31, may have chaperone activity itsel f, preventing heat-induced aggregation of substrate proteins, including α -synuclein. In addition, several lines of evidence suggest that DJ-1 plays a role in human tumorigenesis. First, breast cancer patients have elevated levels of circulating DJ-1 and anti-DJ-1 autoantibodies compared to healthy and non-breast cancer patients. Secondly, DJ-1 protein is increased in primary non-small cell lung carcinoma sa mples. Thirdly, treatment of cells from the human lung cancer cell line NCI-H157 with paclitaxel and MEK inhibitor U0126 leads to a decrease in DJ-1 protein expression.

UniProt: Q99497

Pathways: Intracellular Steroid Hormone Receptor Signaling Pathway, Regulation of Intracellular Steroid Hormone Receptor Signaling, Proton Transport