Park2

Manufactured by Abcam

Sourced in United States

PARK2 is a protein that is involved in the regulation of mitochondrial function and quality control. It plays a role in the process of mitochondrial autophagy, also known as mitophagy, which is the selective degradation of damaged or dysfunctional mitochondria. PARK2 acts as an E3 ubiquitin-protein ligase, attaching ubiquitin molecules to target proteins, which then initiates the mitophagy pathway.

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Lab products found in correlation

Sqstm1 p62, by Abcam (4 mentions) Pink1, by Abcam (4 mentions) Ripa lysis buffer, by Beyotime (3 mentions) Gapdh, by Proteintech (2 mentions) Rapamycin, by Merck Group (1 mentions) Chloroquine, by Merck Group (1 mentions) Carbonyl cyanide 3 chlorophenylhydrazone, by Merck Group (1 mentions) Iba 1, by Abcam (1 mentions) Neuronal nuclei (neun), by Merck Group (1 mentions) Hoechest 33342, by Thermo Fisher Scientific (1 mentions) Lysosensor green dnd 189, by Thermo Fisher Scientific (1 mentions) Mitosox, by Thermo Fisher Scientific (1 mentions) Tom20, by Abcam (1 mentions) 3 methyladenine 3 ma, by Merck Group (1 mentions) Mitotracker green fm, by Thermo Fisher Scientific (1 mentions) Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions) Glial fibrillary acidic protein (gfap), by Santa Cruz Biotechnology (1 mentions) Hrp conjugated goat anti rabbit igg, by Cell Signaling Technology (1 mentions) Hrp conjugated goat anti mouse igg, by Cell Signaling Technology (1 mentions) Trpa1, by Novus Biologicals (1 mentions)

Spelling variants of this product

Anti-PARK2 (4 citations)

5 protocols using park2

Autophagy and Mitophagy Regulation Assay

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Morphine was purchased from Shenyang First Pharmaceutical Factory, Shenyang, China. Rapamycin, 3-methyladenine (3-MA), rotenone, carbonyl cyanide 3-chlorophenylhydrazone (CCCP), and chloroquine (CQ) were purchased from Sigma. MitoSOX, MitoTracker Green FM, LysoSensor Green DND-189, LysoTracker Red DND-99, and Hoechest 33342 were from Life Technologies.
The primary antibodies for immunoblotting analysis were as follows: LC3A/B (CST, 4108), SQSTM1/p62 (CST, 5114), ATG-5 (CST, 12994), BECN1 (CST, 3738), BAX (CST, 2772), BCL2 (CST, 2876), Tom20 (Abcam, ab186734), COX IV (CST, 11967), PARK2 (Abcam, ab15954), PARK2-phospho S65 (Abcam, ab154995), PINK1 (CST, 6946), ATPB (Abcam, ab14730), and ACTB (CST, 4970). The secondary antibodies were from Cell Signaling Technology.
The primary antibodies for immunofluorescence analysis were as follows: SQSTM1/p62 (Abcam, ab91526/56416), NeuN (Millipore, MAB337), GFAP (Santa Cruz, sc-6170), Iba1 (Abcam, ab5076), and PARK2 (Abcam, ab15954). The secondary antibodies were from Jackson ImmunoResearch.

Kong H., Jiang C.Y., Hu L., Teng P., Zhang Y., Pan X.X., Sun X.D, & Liu W.T. (2019). Morphine induces dysfunction of PINK1/Parkin-mediated mitophagy in spinal cord neurons implying involvement in antinociceptive tolerance. Journal of Molecular Cell Biology, 11(12), 1056-1068.

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Mitochondrial Dynamics in Lung Tissues

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Total proteins were extracted from lung tissues of mice using a RIPA Lysis Buffer (Beyotime) and protein concentrations were determined by a BCA assay kit (Beyotime). 30 µg protein per lane was separated using 8–10% SDS PAGE, transferred to a 0.45-µm PVDF membrane, blocked with 5% non-fat milk, and incubated with primary antibodies against DRP1, MFF, OPA1, MFN2 (Cell Signaling Technology, Danvers, MA, USA), PINK1, PARK2, SQSTM1/p62 (Abcam, Cambridge, MA, USA), TRPA1 (Novus Biologicals, Littleton, CO, USA), TRPV1 (Immunoway, Newark, DE, USA ) and GAPDH (Proteintech, Wuhan, Hubei, China) overnight at 4 ℃. Membranes were incubated with the secondary antibodies, HRP-conjugated goat anti-rabbit IgG or HRP-conjugated goat anti-mouse IgG (all 1:3,000, Cell Signaling Technology) for 2 h at room temperature, and the bands were visualized by chemiluminescence.

Li C., Zhang H., Wei L., Liu Q., Xie M., Weng J., Wang X., Chung K.F., Adcock I.M., Chen Y, & Li F. (2022). Role of TRPA1/TRPV1 in acute ozone exposure induced murine model of airway inflammation and bronchial hyperresponsiveness. Journal of Thoracic Disease, 14(7), 2698-2711.

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Protein Expression Profiling of Mitochondrial Dynamics

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By homogenization and lysis in RIPA Lysis Buffer (Beyotime), we extracted total proteins from lung tissues, primary ATII cells, and A549 cells. The protein content was quantified using a BCA kit (Beyotime). Western Blot analysis in lung tissues and cells was performed against DRP1, phosphorylated-DRP1 (p-DRP1) (Ser616), MFF, OPA1, MFN2 (1:1000, Cell Signaling Technology, Danvers, MA, USA), PINK1, PARK2, SQSTM1/p62, LC3b, P16, H2AX (1:1000, Abcam Cambridge, MA, USA), Klotho and GAPDH (1:1000, Proteintech, Wuhan, Hubei, China). Bands were developed by ECL chemiluminescent substrate (Millipore, Billerica, MA, USA).

Li C., Liu Q., Chang Q., Xie M., Weng J., Wang X., Li M., Chen J., Huang Y., Yang X., Wang K., Zhang N., Chung K.F., Adcock I.M., Zhang H, & Li F. (2023). Role of mitochondrial fusion proteins MFN2 and OPA1 on lung cellular senescence in chronic obstructive pulmonary disease. Respiratory Research, 24, 319.

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Immunoblotting Analysis of Mitochondrial Dynamics

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Total protein from A549 cells cultured in 6-well plates and mice lung tissues were respectively homogenized with a RIPA lysis buffer (Beyotime), and protein concentrations were quantified by a Bicinchoninic Acid (BCA) assay kit (Thermo Fisher Scientific, Waltham, MA, USA). 30 µg of protein per lane was separated through 10–15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel and transferred to polyvinylidene fluoride (PVDF) membrane. The membranes were blocked with 5% nonfat milk and incubated with the following primary antibodies: DRP1 (1:1000, Cell Signaling Technology, Danvers, MA,USA), MFF (1:1000, Cell Signaling Technology), MFN2 (1:1000, Cell Signaling Technology), OPA1 (1:1000, Cell Signaling Technology), phosphorylated (phospho) DRP1 (p-DRP1)(1:1000, Cell Signaling Technology), PINK1 (1:1000, Abcam), PARK2 (1:1000, Abcam), SQSTM1/p62 (1:1000, Abcam), LC3 (1:500, Abcam), MLKL (1:1000, Affinity Biosciences, Liyang, Jiangsu, China), RIPK1 (1:1000, Cell Signaling Technology), RIPK3 (1:1000, Cell Signaling Technology), overnight at 4 °C. Membranes were then incubated with a HRP-conjugated anti-rabbit secondary antibody (Cell Signaling Technology,USA) and then visualized by chemiluminescent detection.

Liu Q., Weng J., Li C., Feng Y., Xie M., Wang X., Chang Q., Li M., Chung K.F., Adcock I.M., Huang Y., Zhang H, & Li F. (2023). Attenuation of PM2.5-induced alveolar epithelial cells and lung injury through regulation of mitochondrial fission and fusion. Particle and Fibre Toxicology, 20, 28.

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Protein Expression in Kidney Samples

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A piece from flash frozen kidneys were homogenized in RIPA buffer containing cocktail protease and phosphatase inhibitors. Kidneys representative of mean serum creatinine for each group were used for Western blot samples. Total protein content was measured by the BCA assay. 20 μg total protein was loaded into SDS-PAGE gels and immunoblots were performed as previously described (39) . Antibodies used include peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α; 1:1,000; Cat. No. ab54481; Abcam); mitochondrial transcription factor A (Tfam; 1:1,000; developed in our laboratory); Pink1 (1:500; cat. no. ab23707; Abcam); PARK2 (1:500; Cat. No. sc-32282; Santa Cruz Biotechnology); light chain 3B protein (LC3I/II) tubulin (1:1,000; Cat. No. T-5168; Sigma-Aldrich), or mitochondrial porin (1:500; Cat. No. sc-8829), Drp1 (Santa Cruz Biotechnology, Dallas, TX), sirtuin 3 (#5490S, Cell Signaling Technologies, Danvers, MA).

Suliman H.B., Ma Q., Zhang Z., Ren J., Morris B.T., Crowley S.D., Ulloa L., & Privratsky J.R. (2020). Annexin A1 tripeptide mimetic increases sirtuin-3 to augment mitochondrial function and limit ischemic kidney injury.

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