Product Datasheet. PINK1 Antibody BC SS. Unit Size: ml

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1 Product Datasheet PINK1 Antibody BC SS Unit Size: ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Reviews: 10 Publications: 117 Protocols, Publications, Related Products, Reviews, Research Tools and Images at: Updated 2/3/2019 v.20.1 Earn rewards for product reviews and publications. Submit a publication at Submit a review at

2 BC SS PINK1 Antibody Product Information Unit Size Concentration Storage Clonality Preservative Isotype Purity Buffer Target Molecular Weight ml 1.0 mg/ml Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. Polyclonal 0.02% Sodium Azide IgG Immunogen affinity purified PBS 63 kda Page 1 of 6 v.20.1 Updated 2/3/2019 Product Description Host Rabbit Gene ID Gene Symbol Species PINK1 Human, Mouse, Rat, Drosophila, Rabbit Reactivity Notes Drosophila reactivity reported in scientific literature (PMID: ). Immunogen A synthetic peptide made to the human PINK1 protein sequence (between residues ). [Swiss-Prot Q9BXM7] Product Application Details Applications Recommended Dilutions Application Notes Western Blot, Immunocytochemistry/Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Peptide ELISA Western Blot 1:500-1:2000, Immunohistochemistry, Immunocytochemistry/Immunofluorescence 1:50-1:200, Immunoprecipitation, Immunohistochemistry-Paraffin, Peptide ELISA 1:100-1:2000 This PINK1 antibody can be used for ICC, Peptide ELISA and Western blot, where specific bands are seen at 48, 55 and 63 kda. In WB, this antibody has been used in valinomycin and CCCP treated HeLa whole cell lysate. Use in IP reported in scientific literature (PMID ) Use in paraffin-sections reported in scientific literature (PMID ). In Western blot it is highly recommended to use 1-5% w/v BSA in TBS with 0.1% Tween-20 for all incubations.

3 Images Western Blot: PINK1 Antibody [BC ] - Analysis of PINK1 in HeLa whole cell lysate with and without treatment of 10 um CCCP. Image courtesy of an anonymous customer review. Page 2 of 6 v.20.1 Updated 2/3/2019 Immunocytochemistry/Immunofluorescence: PINK1 Antibody [BC ] - Immunocytochemistry of PINK1 antibody (BC Lot G). HeLa cells were treated with valinomycin (1 um for 24h) prior to being fixed in 10% buffered formalin for 10 min and permeabilized in 0.1% Triton X-100 in PBS for 10 min. Cells were incubated with BC at 20 ug/ml for 1h at room temperature, washed 3x in PBS and incubated with Alexa-Fluor488 anti-rabbit secondary antibody. PINK1 (Green) was detected at the mitochondria. Tubulin (Red) was detected using an antitubulin antibody with an anti-mouse DyLight 550 secondary antibody. DNA (Blue) was counterstained with DAPI. Note: mitochondria staining might not be easily observed without treatment with valinomycin or CCCP. Immunocytochemistry/Immunofluorescence: PINK1 Antibody [BC ] - HeLa cells were treated with valinomycin (1 um for 24h) prior to being fixed in 10% buffered formalin for 10 min and permeabilized in 0.1% triton X-100 in PBS for 10 min. Cells were incubated with BC at 20 ug/ml for 1h at room temperature, washed 3x in PBS and incubated with Alexa-Fluor488 anti-rabbit secondary antibody. PINK1 (Green) was detected at the mitochondria. Tubulin (Red) was detected using NB with an anti-mouse DyLight 550 secondary antibody. DNA (Blue) was counterstained with DAPI. Note: mitochondrial staining may only be visible after treatment with valinomycin or CCCP. Western Blot: PINK1 Antibody [BC ] - Analysis of PINK1 in mouse liver and hypatocytes using PINK1 antibody. Image from verified customer review.

4 Western Blot: PINK1 Antibody [BC ] - WB analysis of lysates derived from htert-rpe1 cells transfected with non-targeting (NT1) or PINK1 targeting sirna for 72 h. 20 ug were loaded. Image from verified customer review. Page 3 of 6 v.20.1 Updated 2/3/2019 Immunohistochemistry-Paraffin: PINK1 Antibody [BC ] - Rabbit heart tissue. Image from verified customer review. Western Blot: PINK1 Antibody [BC ] - Western blot image of PINK1 antibody (BC ) in multiple cells lines. Human HeLa (lane 1), Mouse NIH-3T3 (lane 2), L929 (lane 3) and Rat PC12 (lane 4) whole cell protein were separated by SDS-PAGE on a 7.5% polyacrylamide gel. Protein was transferred to PVDF membrane and probed with 2 ug/ml BC in 1% BSA and detected with an HRP-conjugated anti-rabbit secondary antibody using chemiluminescence. PINK1 was detected at approximately 60 kda (arrowhead). Western Blot: PINK1 Antibody [BC ] - Whole cell protein from HeLa cells treated without or with valinomycin (1 um for 24h) as indicated were separated by SDS-PAGE on a 7.5% polyacrylamide gel. Protein was transferred to PVDF membrane and probed with 1.0 ug/ml BC in 1% BSA and detected with an HRP-conjugated anti-rabbit secondary antibody using chemiluminescence. PINK1 was detected at approximately 60 kda in the treated sample(arrowhead).

5 Publications McWilliams TG, Barini E, Pohjolan-Pirhonen R et al. Phosphorylation of Parkin at serine 65 is essential for its activation in vivo. Open Biol. Nov :00AM [PMID: ] (WB, Mouse) Kim HJ, Joe Y, Rah SY et al. Carbon monoxide-induced TFEB nuclear translocation enhances mitophagy/mitochondrial biogenesis in hepatocytes and ameliorates inflammatory liver injury. Cell Death Dis. Oct :00AM [PMID: ] (WB, Mouse) Civiletto G, Dogan SA, Cerutti R et al. Rapamycin rescues mitochondrial myopathy via coordinated activation of autophagy and lysosomal biogenesis. EMBO Mol Med. Oct :00AM [PMID: ] (WB, Mouse) Araya J, Tsubouchi k, Sato N et al. PRkN-regulated mitophagy and cellular senescence during COPD pathogenesis. Autophagy. Oct :00AM [PMID: ] (WB, Human) Walsh TG, van den Bosch MTJ, Lewis KE et al. Loss of the mitochondrial kinase PINK1 does not alter platelet function. Sci Rep Sep :00AM [PMID: ] (WB, Mouse) Sun X, Duan Y, Qin C et al. Distinct multilevel misregulations of Parkin and PINK1 revealed in cell and animal models of TDP-43 proteinopathy. Cell Death Dis Sep :00AM [PMID: ] (WB, Drosophila) Basso V, Marchesan E, Peggion C et al. Regulation of Endoplasmic Reticulum-Mitochondria contacts by Parkin via Mfn2. Pharmacol. Res. Sep :00AM [PMID: ] (WB, Mouse) Lazarou Michael, Narendra Derek P, Jin Seok Min et al. PINK1 drives Parkin self-association and HECT-like E3 activity upstream of mitochondrial binding. J Cell Biol 2013 [PMID: ] (WB, Human) Thomas HE, Zhang Y, Stefely JA et al. Mitochondrial Complex I Activity Is Required for Maximal Autophagy Cell Rep Aug :00AM [PMID: ] (WB, Human) Marcassa E, Kallinos A, Jardine J et al. Dual role of USP30 in controlling basal pexophagy and mitophagy EMBO Rep. Jun :00AM [PMID: ] (Human) Sun Y, Yao X, Zhang QJ et al. Beclin-1-Dependent Autophagy Protects the Heart During Sepsis Circulation May :00AM [PMID: ] (Mouse) Goiran T, Duplan E, Rouland L et al. Nuclear p53-mediated repression of autophagy involves PINK1 transcriptional down-regulation Cell Death Differ. May :00AM [PMID: ] (WB, Mouse) More publications at Page 4 of 6 v.20.1 Updated 2/3/2019

6 Procedures Western Blot Protocol for PINK1 Antibody (BC ) Western Blot Protocol Page 5 of 6 v.20.1 Updated 2/3/ Perform SDS-PAGE on samples to be analyzed, loading ug of total protein per lane. 2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus. 3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate. 4. Rinse the blot of the protein stain. 5. Block the membrane using 5% BSA for at least 1 hour. 6. Dilute anti-pink1 primary antibody in 1% BSA and incubate 1 hour at room temperature. 7. Wash the membrane in wash buffer three times for 10 minutes each. 8. Incubate in diluted HRP-conjugated Rabbit secondary antibody in 1% BSA (as per manufacturers instructions) and incubate 1 hour at room temperature. 9. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background). 10. Apply the detection reagent of choice in accordance with the manufacturers instructions.

7 Novus Biologicals USA E. Briarwood Avenue Centennial, CO USA Phone: Toll Free: Fax: Novus Biologicals Canada 461 North Service Road West, Unit B37 Oakville, ON L6M 2V5 Canada Phone: Toll Free: Fax: Novus Biologicals Europe 19 Barton Lane Abingdon Science Park Abingdon, OX14 3NB, United Kingdom Phone: (44) (0) Free Phone: Fax: (44) (0) General Contact Information Technical Support: Orders: General: Limitations This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt. For more information on our 100% guarantee, please visit Earn gift cards/discounts by submitting a review: Earn gift cards/discounts by submitting a publication using this product: