PINK1 Antibody - BSA Free

Novus Biologicals | Catalog # NB600-973

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all PINK1 Primary Antibodies »

Product Specifications

Immunogen

PINK1 antibody was developed using a synthetic peptide from human PINK1, corresponding to amino acids 484-504 Human:LVRALLQREASKRPSARVAAN
Mouse:LVRsLLQREASKRPSARVAAN

Localization

Mitochondrial

Specificity

PINK1 polyclonal antibody recognizes primarily the full length protein at about 66 kDa in human, mouse, and rat tissues. In addition, a truncated form of the protein at about 33 kDa is also detected.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Theoretical MW

62.7 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for PINK1 Antibody - BSA Free

Western Blot: PINK1 Antibody [NB600-973]

Western Blot: PINK1 Antibody [NB600-973]

Western Blot: PINK1 Antibody [NB600-973] - Lane 1: The primary band with the observed molecular weight of 66 kDa was detected in mouse liver (30ug). Also, a truncated form of the protein at about 33 kDa was detected.
Immunocytochemistry/ Immunofluorescence: PINK1 Antibody [NB600-973]

Immunocytochemistry/ Immunofluorescence: PINK1 Antibody [NB600-973]

PINK1-Antibody-Immunocytochemistry-Immunofluorescence-NB600-973-img0003.jpg
Immunohistochemistry-Paraffin: PINK1 Antibody [NB600-973]

Immunohistochemistry-Paraffin: PINK1 Antibody [NB600-973]

Immunohistochemistry-Paraffin: PINK1 Antibody [NB600-973] - Staining in human liver tissue using a 1:200 dilution.
PINK1 Antibody - BSA Free

Western Blot: PINK1 Antibody - BSA Free [NB600-973] -

Overexpression of PINK1 in SCs of ETRs indicating mitochondrial damage. (A) IHC of PINK1. Note the perinuclear localization of PINK1 (arrows). The broken arrows indicate SC nuclei. The inset shows magnified SC nuclei; (B,C) Western blot analysis of PINK1. The relative expression level for protein was normalized to actin and expressed as fold change relative to the control (n = 3). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30621351), licensed under a CC-BY license. Not internally tested by Novus Biologicals.
PINK1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: PINK1 Antibody - BSA Free [NB600-973] -

LRRK2 accumulates in globules in alpha S tg mice. (a and b) Double immunofluorescence for alpha S with parkin, PINK1, DJ-1, LRRK2, or negative control (the immunopeptide-preabsorbed anti-LRRK2 antibody) in alpha S tg mice (a) and P123H beta S tg mice (b). Note that alpha S-globules were immunopositive for LRRK2 (~79%, n = 22), whereas P123H beta S globules were negative for LRRK2. Representative images are shown for the thalamus ( alpha S) and basal ganglia (P123H beta S). Scale bar = 5 μm for all panels. (c) Triple immunofluorescence for alpha S, LRRK2 and Rab5B for basal ganglia in alpha S tg mice. LRRK2 and Rab5B were colocalized in axon terminal (arrow), but were not colocalized in the alpha S-globule (arrowhead) Scale bar = 10 μm for all panels. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/23013868), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Applications for PINK1 Antibody - BSA Free

Application
Recommended Usage

Immunohistochemistry

4ug/ml

Immunohistochemistry-Paraffin

4ug/ml

Western Blot

1:150
Application Notes
IHC-Frozen reactivity reported in ( PMID: 23013868), IHC-P reactivity reported in ( PMID: 26935412).

Formulation, Preparation, and Storage

Purification

Peptide affinity purified

Formulation

PBS, pH 7.2, containing 50% glycerol

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: PINK1

Phosphatase and Tensin Homolog (PTEN) is a tumor suppressor which acts as an antagonist to phosphatidylinositol 3-kinase (PI3K) signaling. PTEN exerts enzymatic activity as a phosphatidylinositol-3,4,5-trisphosphate (PIP3) phosphatase, opposing PI3K activity by reducing availability of PIP3 to proliferating cells. Loss of PTEN function leads to elevated PIP3 and increased activation of PI3K/AKT signaling in many types of cancer.

PINK1 (PTEN induced putative kinase 1) protein contains a N-terminal mitochondrial targeting sequence, putative transmembrane helix, linker region, serine (Ser65)/threonine (Thr257) kinase domain and C-terminal segment. PINK1 is translated in the cytosol, then translocated to the outer mitochondrial membrane where it is rapidly cleaved and degraded as a part of normal mitochondrial function. In damaged (depolarized) mitochondria, PINK1 becomes stabilized and accumulates, resulting in the subsequent phosphorylation of numerous proteins on the mitochondrial surface.

When PINK1 is imported into the cell, mitochondrial processing peptidase, presenilin-associated rhomboid-like protease and AFG3L2 cleave PINK1 and tag it for the ubiquitin-proteasome pathway, keeping low PINK1 protein expression at basal conditions (1,2). Accumulation of PINK1 in mitochondria indicate damage. PINK1 maintains mitochondrial function/integrity, provides protection against mitochondrial dysfunction during cellular stress, and is involved in the clearance of damaged mitochondria via selective autophagy (mitophagy) (3). PINK1 has a theoretical molecular weight of 63 kDa and undergoes proteolytic processing to generate at least two cleaved forms (55 kDa and 42 kDa).

Ultimately PARK2 (E3 Ubiquitin Ligase Parkin) is recruited to the damaged mitochondria where it is activated by 1) PINK-mediated phosphorylation of PARK2 at serine 65, and 2) PARK2 interaction with phosphorylated ubiquitin (also phosphorylated by PINK1 on serine 65) (4,5). There is a strong interplay between Parkin and PINK1, where loss-of-function of human PINK1 results in mitochondrial pathology and can be rescued by Parkin (2,4,5). Mutations in either Parkin or PINK1 alter mitochondrial turnover, resulting in the accumulation of defective mitochondria and, ultimately, neurodegeneration in Parkinson's disease. Mutations in the PINK1 gene located within the PARK6 locus on chromosome 1p35-p36 have been identified in patients with early-onset Parkinson's disease (6).

References

1.Rasool, S., Soya, N., Truong, L., Croteau, N., Lukacs, G. L., & Trempe, J. F. (2018). PINK1 autophosphorylation is required for ubiquitin recognition. EMBO Rep, 19(4). doi:10.15252/embr.201744981

2.Shiba-Fukushima, K., Arano, T., Matsumoto, G., Inoshita, T., Yoshida, S., Ishihama, Y.,... Imai, Y. (2014). Phosphorylation of mitochondrial polyubiquitin by PINK1 promotes Parkin mitochondrial tethering. PLoS Genet, 10(12), e1004861. doi:10.1371/journal.pgen.1004861

3.Vives-Bauza, C., Zhou, C., Huang, Y., Cui, M., de Vries, R. L., Kim, J.,... Przedborski, S. (2010). PINK1-dependent recruitment of Parkin to mitochondria in mitophagy. Proc Natl Acad Sci U S A, 107(1), 378-383. doi:10.1073/pnas.0911187107

4.McWilliams, T. G., Barini, E., Pohjolan-Pirhonen, R., Brooks, S. P., Singh, F., Burel, S.,... Muqit, M. M. K. (2018). Phosphorylation of Parkin at serine 65 is essential for its activation in vivo. Open Biol, 8(11). doi:10.1098/rsob.180108

5.Exner, N., Treske, B., Paquet, D., Holmstrom, K., Schiesling, C., Gispert, S.,... Haass, C. (2007). Loss-of-function of human PINK1 results in mitochondrial pathology and can be rescued by parkin. J Neurosci, 27(45), 12413-12418. doi:10.1523/jneurosci.0719-07.2007

6.Valente, E. M., Bentivoglio, A. R., Dixon, P. H., Ferraris, A., Ialongo, T., Frontali, M.,... Wood, N. W. (2001). Localization of a novel locus for autosomal recessive early-onset parkinsonism, PARK6, on human chromosome 1p35-p36. Am J Hum Genet, 68(4), 895-900. doi:10.1086/319522

Long Name

PTEN-induced Putative Kinase 1

Alternate Names

BRPK, PARK6

Entrez Gene IDs

65018 (Human); 68943 (Mouse); 298575 (Rat)

Gene Symbol

PINK1

UniProt

Q9BXM7 (Human)

Additional PINK1 Products

Product Documents for PINK1 Antibody - BSA Free

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Product Specific Notices for PINK1 Antibody - BSA Free

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.

Citations for PINK1 Antibody - BSA Free

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for PINK1 Antibody - BSA Free

Showing  1 - 2 of 2 FAQs Showing All

  • Q: I bought PINK1 Antibody (NB600-973) from your company. According to data sheet, PINK1 polyclonal antibody recognizes primarily the full length protein at about 66 kDa in addition to about truncated 33 kDa based on western blot. Please could you tell me how immunohistochemistry could differentiate the two forms of protein? So, please could you tell me the subcelluar localization of PINK1? Could be nuclear under certain conditions? I know that PINK1 is mitochondrial protein, however, is the pattern of this protein is diffuse cytoplasmic or punctuate?

    A: In answer to your question, because the antibody binds to both forms, and Western blot distinguishes between the two by separating by size (gel electrophoresis), which immunohistochemistry does not do, IHC would not differentiate between the full-length and truncated forms. The following journal article discusses the subcellular and submitochondrial localization of PINK1, so hopefully will be of use to you: The kinase domain of mitochondrial PINK1 faces the cytoplasm (https://www.pnas.org/doi/pdf/10.1073/pnas.0802814105)

  • Q: I’m performing western blots on a neuronal cell line using RIPA buffer, sadly, we are only able to detect the 30 kDa isoform.  Do you have any suggestions for detecting the 66 kDa isoform?

    A: Hello and thank you for contacting Novus Biological’s tech line. I would recommend that you use an antibody that targets the N-terminal position of the protein between 78 to 110. Antibody NB100-493 targets the topological mitochondrial intermembrane domain, so there may be a better opportunity of detecting the pre-processed protein.  Also, I would suggest that you use an overexpression PINK1 lysate as a positive control.

  • Q: I bought PINK1 Antibody (NB600-973) from your company. According to data sheet, PINK1 polyclonal antibody recognizes primarily the full length protein at about 66 kDa in addition to about truncated 33 kDa based on western blot. Please could you tell me how immunohistochemistry could differentiate the two forms of protein? So, please could you tell me the subcelluar localization of PINK1? Could be nuclear under certain conditions? I know that PINK1 is mitochondrial protein, however, is the pattern of this protein is diffuse cytoplasmic or punctuate?

    A: In answer to your question, because the antibody binds to both forms, and Western blot distinguishes between the two by separating by size (gel electrophoresis), which immunohistochemistry does not do, IHC would not differentiate between the full-length and truncated forms. The following journal article discusses the subcellular and submitochondrial localization of PINK1, so hopefully will be of use to you: The kinase domain of mitochondrial PINK1 faces the cytoplasm (https://www.pnas.org/doi/pdf/10.1073/pnas.0802814105)

  • Q: I’m performing western blots on a neuronal cell line using RIPA buffer, sadly, we are only able to detect the 30 kDa isoform.  Do you have any suggestions for detecting the 66 kDa isoform?

    A: Hello and thank you for contacting Novus Biological’s tech line. I would recommend that you use an antibody that targets the N-terminal position of the protein between 78 to 110. Antibody NB100-493 targets the topological mitochondrial intermembrane domain, so there may be a better opportunity of detecting the pre-processed protein.  Also, I would suggest that you use an overexpression PINK1 lysate as a positive control.

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