PINK1 Antibody - BSA Free

Images

 
Biological Strategies: Knockdown Validated: PINK1 Antibody [BC100-494] - HEK293 cells were co-transfected with PINK1 siRNA (#1 or #2) or scrambled siRNA (scrambled) and untagged wild-type (WT) or Ser65Ala (S65A) ...read more
Electron Microscopy: PINK1 Antibody [BC100-494] - Representative TEM picture. The red and green arrows indicate PINK and PARKIN immunogold particles in the dKO RPE samples, respectively (e). ML, melanosome. Scale = 0.2 ...read more
Biological Strategies: Western Blot: PINK1 Antibody [BC100-494] - Pathogenic mutants of Parkin are subjected to Ser65 phosphorylation. Phos-tag Western blotting for Parkin and Western blotting for PINK1 were ...read more
Immunocytochemistry/ Immunofluorescence: PINK1 Antibody [BC100-494] - Immunocytochemistry of PINK1 antibody (BC100-494 Lot G). HeLa cells were treated with valinomycin (1 uM for 24h) prior to being fixed in 10% buffered ...read more
Western Blot: PINK1 Antibody [BC100-494] - 35 ug Neuro2A whole cell lysate separated on 8% PAGE and stained for PINK1 (1:1000 in 5% milk powder in TBST; secondary antibody AP at 1:5000). WB image submitted by a verified ...read more
Immunohistochemistry-Paraffin: PINK1 Antibody [BC100-494] - Rabbit heart tissue. IHC-P image submitted by a verified customer review.
Western Blot: PINK1 Antibody [BC100-494] - Western blot image of PINK1 antibody (BC100-494) in multiple cells lines. Human HeLa (lane 1), Mouse NIH-3T3 (lane 2), L929 (lane 3) and Rat PC12 (lane 4) whole cell protein ...read more
Western Blot: PINK1 Antibody [BC100-494] - Analysis of PINK1 in mouse liver and hypatocytes using PINK1 antibody. Observed molecular weight ~55 kDa. WB image submitted by a verified customer review.
Western Blot: PINK1 Antibody [BC100-494] - Alteration of mitochondria and PD associated proteins in SH-SY5Y cells with telomere removal by CRISPR-Cas9. Representative Western blot of PGC-1alpha, NRF1, and PINK1 in ...read more
Genetic Strategies: Knockout Validated: PINK1 Antibody [BC100-494] - Characterisation of mitochondrial content and function in PINK1-/- platelets. Cropped immunoblots showing PINK1 expression in 10 uM CCCP (6 ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Dr, RbSpecies Glossary
Applications WB, EM, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PAGE, PEP-ELISA, KD, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml

PINK1 Antibody - BSA Free Summary

Immunogen
PINK1 antibody was developed using a synthetic peptide made to the human PINK1 protein sequence (between residues 175-250). [Swiss-Prot Q9BXM7]
Localization
Localizes mostly in mitochondrion and the 2 proteolytic processed fragments (Topological domain 111-581) of 55 kDa and 48 kDa localize mainly in cytosol.
Specificity
Human PINK1 Antibody will be reactive to isoform 2.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
PINK1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Electron Microscopy
  • Immunoblotting
  • Immunocytochemistry/Immunofluorescence 1:50 - 1:200
  • Immunohistochemistry
  • Immunohistochemistry-Frozen
  • Immunohistochemistry-Paraffin
  • Immunoprecipitation
  • Knockdown Validated
  • Knockout Validated
  • Peptide ELISA 1:100 - 1:2000
  • SDS-Page
  • Western Blot 1:500 - 1:2000
Application Notes
NOTE: It's recomended to use 1-5% w/v BSA in TBS with 0.1% Tween-20 for all incubations in WB. 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: 22078885) Use in paraffin-sections reported in scientific literature (PMID: 25083992). Use in Immunoblotting reported in multiple pieces of scientific literature. Use in SDS-PAGE reported in scientific literature (PMID: 27846363). Use in Knockout Validated reported in scientific literature (PMID: 31066324). Use in Immunohistochemistry-Frozen reported in scientific literature (PMID: 31908016).
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.
Control Peptide
PINK1 Protein (BC100-494PEP)
Reviewed Applications
Read 13 Reviews rated 3.9
using
BC100-494 in the following applications:

Publications
Read Publications using
BC100-494 in the following applications:

  • 29 publications
  • IHC
    4 publications
  • 1 publication
  • 3 publications
  • IP
    11 publications
  • KD
    3 publications
  • KO
    1 publication
  • 1 publication
  • WB
    142 publications

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:33775690). All species in which poly(GP) peptides are synthesized. Human reactivity reported in multiple pieces of scientific literature.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for PINK1 Antibody - BSA Free

  • Anti-PINK1 Antibody
  • BRPK Antibody
  • BRPK
  • EC 2.7.11.1
  • FLJ27236
  • Mitochondrial Antibody
  • PARK6
  • Parkinson disease (autosomal recessive) 6
  • PINK1 Antibody
  • PINK1 mouse
  • PINK1 polyclonal
  • PINK1
  • protein kinase BRPK
  • PTEN Induced Kinase 1
  • PTEN induced putative kinase 1
  • PTEN-induced putative kinase protein 1
  • serine/threonine-protein kinase PINK1, mitochondrial

Background

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

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.

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BC100-494
Species: Hu, Mu, Rt, Dr, Rb
Applications: WB, EM, IB, ICC/IF, IHC, IHC-Fr, IHC-P, IP, PAGE, PEP-ELISA, KD, KO

Publications for PINK1 Antibody (BC100-494)(205)

We have publications tested in 4 confirmed species: Human, Mouse, Rat, Drosophila.

We have publications tested in 9 applications: ICC/IF, IHC, IHC-F, IHC-P, IP, KD, KO, Knockdown, WB.


Filter By Application
ICC/IF
(29)
IHC
(4)
IHC-F
(1)
IHC-P
(3)
IP
(11)
KD
(3)
KO
(1)
Knockdown
(1)
WB
(142)
All Applications
Filter By Species
Human
(99)
Mouse
(53)
Rat
(8)
Drosophila
(1)
All Species
Showing Publications 1 - 10 of 205. Show All 205 Publications.
Publications using BC100-494 Applications Species
Gillmore T Mitochondrial Dynamics in Mesenchymal Cells in Placental Development and Preeclampsia Cell Death Dis Feb 27 2022 [PMID: 35220394]
Yang L, Tao Y, Luo L et al. Dengzhan Xixin injection derived from a traditional Chinese herb Erigeron breviscapus ameliorates cerebral ischemia/reperfusion injury in rats via modulation of mitophagy and mitochondrial apoptosis Journal of ethnopharmacology Jan 12 2022 [PMID: 35032588] (IHC, Rat) IHC Rat
Gillmore T Mitochondrial Dynamics in Mesenchymal Cells in Placental Development and Preeclampsia Thesis
Zhang Y, Fang Q, Wang H et al. Increased mitophagy protects cochlear hair cells from aminoglycoside-induced damage Autophagy Apr 26 2022 [PMID: 35471096] (WB, Mouse) WB Mouse
Vianello C, Cocetta V, Catanzaro D et al. Cisplatin resistance can be curtailed by blunting Bnip3-mediated mitochondrial autophagy Cell death & disease Apr 22 2022 [PMID: 35459212] (WB, Human) WB Human
Sonn S, Song E, Seo S et al. Peroxiredoxin 3 deficiency induces cardiac hypertrophy and dysfunction by impaired mitochondrial quality control Redox Biology May 1 2022 [PMID: 35248828] (WB, Mouse) WB Mouse
Harbauer AB, Hees JT, Wanderoy S et al. Neuronal mitochondria transport Pink1 mRNA via synaptojanin 2 to support local mitophagy Neuron Feb 19 2022 [PMID: 35216662]
Li J, Lai M, Zhang X et al. PINK1-parkin-mediated neuronal mitophagy deficiency in prion disease Cell death & disease Feb 18 2022 [PMID: 35184140] (WB, Mouse) WB Mouse
Jiang XJ, Wu YQ, Ma R et al. PINK1 Alleviates Cognitive Impairments via Attenuating Pathological Tau Aggregation in a Mouse Model of Tauopathy Frontiers in cell and developmental biology Jan 4 2022 [PMID: 35059394] (WB) WB
Xing Y, Wei X, Liu Y et al. Autophagy inhibition mediated by MCOLN1/TRPML1 suppresses cancer metastasis via regulating a ROS-driven TP53/p53 pathway Autophagy Dec 8 2021 [PMID: 34878954] (WB) WB
Show All 205 Publications.

Reviews for PINK1 Antibody (BC100-494) (13) 3.913

Average Rating: 3.9
(Based on 13 reviews)
We have 13 reviews tested in 4 species: Human, Mouse, Human, Mouse, Rat, Rabbit.

Reviews using BC100-494:
Filter by Applications
WB
(11)
IF-P
(1)
ELISA
(1)
All Applications
Filter by Species
Human
(7)
Mouse
(4)
Human, Mouse, Rat
(1)
Rabbit
(1)
All Species
Images Ratings Applications Species Date Details
Western Blot PINK1 BC100-494
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5
reviewed by:
Verified Customer
WB Human 11/08/2021
View

Summary

ApplicationWestern Blot
Sample Testedhuman macrophage
SpeciesHuman
LotK-4
Western Blot PINK1 BC100-494
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5
reviewed by:
Verified Customer
WB Mouse 01/09/2021
View

Summary

ApplicationWestern Blot
Sample TestedMouse Neuro2A whole cell lysate
SpeciesMouse
Loto
Western Blot PINK1 BC100-494
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3
reviewed by:
Muhammad Yousuf
WB Mouse 03/23/2019
View

Summary

ApplicationWestern Blot
Sample Testedmouse spinal cord
SpeciesMouse
LotG-1

Comments

CommentsBlocked with 5% BSA in PSB-T. Primary antibody at 1:1000 overnight at 4 degrees followed by secondary antibody at 1:10,000 for 1 hour at room temperature.
Western Blot PINK1 BC100-494
Enlarge
1
reviewed by:
Shinobu Miyazaki
WB Mouse 12/21/2018
View

Summary

ApplicationWestern Blot
Sample Testedmouse vascular smooth muscle cell
SpeciesMouse
LotM
Other PINK1 BC100-494
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4
reviewed by:
Verified Customer
IF-P Rabbit 04/24/2018
View

Summary

ApplicationOther
Sample TestedHeart tissue
SpeciesRabbit
LotG-1
Western Blot PINK1 BC100-494
Enlarge
4
reviewed by:
Verified Customer
WB Human 02/05/2018
View

Summary

ApplicationWestern Blot
Sample TestedhTERT RPE-1 cell line from ATCC,IMCD3, hTertRPE1, HDF, SHSY5Y whole cell lysates,U2OS cell line
SpeciesHuman
LotK-5
  5
reviewed by:
Pradeepkiran Jangampalli Adi
ELISA Human, Mouse, Rat 01/29/2018
View

Summary

ApplicationELISA
Sample TestedELISA Sample Tested,mouse corneal whole-mount immunofluorescence staining
SpeciesHuman, Mouse, Rat
LotAB_10127658
Western Blot PINK1 BC100-494
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5
reviewed by:
Jie Zhuang
WB Human 01/20/2017
View

Summary

ApplicationWestern Blot
Sample TestedHela whole cell lysate
SpeciesHuman
Western Blot PINK1 BC100-494
Enlarge
4
reviewed by:
Liyong Zhang
WB Mouse 08/06/2016
View

Summary

ApplicationWestern Blot
SpeciesMouse
LotPO number: E0513646
Western Blot PINK1 BC100-494
Enlarge
4
reviewed by:
Christian Munch
WB Human 03/31/2016
View

Summary

ApplicationWestern Blot
Sample TestedHela whole cell lysate
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Human 07/05/2011
View

Summary

ApplicationWestern Blot
Sample TestedHela whole cell lysate, Sample Amount: 60ug
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Human 11/10/2010
View

Summary

ApplicationWestern Blot
Sample TestedHeLa whole cell lysate, Sample Amount: 20ug
SpeciesHuman
  3
reviewed by:
Verified Customer
WB Human 05/17/2010
View

Summary

ApplicationWestern Blot
Sample TestedHela whole cell lysate
SpeciesHuman
LotA32

Product General Protocols

View specific protocols for PINK1 Antibody (BC100-494): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for PINK1 Antibody (BC100-494). (Showing 1 - 4 of 4 FAQs).

  1. We recently purchased BC100-494 and the HEK293 cell extract to serve as a positive control. The most prominent band I got is approximately 120 kDa. However, after the cells were treated with DMSO, the most intense band became approximately 60 and 50 kDa (like the datasheet). Since there are some references that mention that PINK1 can form dimer or associate with other mitochondrial proteins, I am wondering if the reason of 120 kDa band is due to the strong binding of protein complex?
    • It is most likely being picked up also at 120 kDa due to associations with other proteins. Our lab has not characterized this higher molecular weight band, but this seems like a likely explanation based on some publications I have found. One which you might find useful is The Mitochondrial Fusion-Promoting Factor Mitofusin Is a Substrate of the PINK1/Parkin Pathway. I recommend trying a negative control competition assay with BC100-494PEP. This blocking peptide can help you determine if this 120 kDa band is specific. You can find a protocol for this procedure that we recommend at the following link, peptide competition protocol.
  2. We would like to check toxicity of BC100-494. Could you inform us of the concentration of sodium azide?
    • This product contains 0.05% NaN3 as the preservative.
  3. I ordered PINK1 antibody(BC100-494) at about six months ago. After it shipped to my country, it has been stored at -20 degrees Celsius in the agent company ever since. I recently called the agent to deliver it to me, and that's when I found this product has been wrongly stored according to the datasheet. So what I am worrying now is that does this influence the efficiency of this antibody? If it does, how much damage has been caused? What should I do to reduce the bad effects? Should I store this antibody by small packages?
    • We recommend that BC100-494 is stored at 4C and that it should not be frozen. Unfortunately we have no testing data regarding the use of the antibody following -20C storage, and as such I cannot guarantee that it will perform optimally now that it has been frozen. I suggest that you try using the antibody as usual, but perhaps at a higher (more concentrated) dilution than recommended in case its potency has been lost during the freezing process. I would store it at 4C from now on, and not freeze it again once you have thawed it, since repeated freeze-that cycles can be detrimental to performance.
  4. Have you tried PINK1 antibody for immunoblotting in HEK293FT cells?
    • We have not performed any testing of this antibody in HEK293FT cells specifically. However, PINK1 is a widely expressed protein, and I would expect it to be present in that particular cell line (it is expressed in HEK293); however, you will most likely want to run this along side controls to confirm.

Secondary Antibodies

 

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Bioinformatics Tool for PINK1 Antibody (BC100-494)

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Blogs on PINK1. Showing 1-10 of 14 blog posts - Show all blog posts.

Understanding Mitophagy Mechanisms: Canonical PINK1/Parkin, LC3-Dependent Piecemeal, and LC3-Independent Mitochondrial Derived Vesicles
By Christina Towers, PhD What is Mitophagy?The selective degradation of mitochondria via double membrane autophagosome vesicles is called mitophagy. Damaged mitochondria can generate harmful amounts of reactive ox...  Read full blog post.

New Players in the Mitophagy Game
By Christina Towers, PhD Mitochondrial turn over via the lysosome, otherwise known as mitophagy, involves engulfment of mitochondria into double membrane autophagosomes and subsequent fusion with lysosomes. Much is al...  Read full blog post.

Losing memory: Toxicity from mutant APP and amyloid beta explain the hippocampal neuronal damage in Alzheimer's disease
 By Jamshed Arslan Pharm.D.  Alzheimer's disease (AD) is an irreversible brain disorder that destroys memory and thinking skills. The telltale signs of AD brains are extracellular deposits of amy...  Read full blog post.

There's an autophagy for that!
By Christina Towers, PhDA critical mechanism that cells use to generate nutrients and fuel metabolism is through a process called autophagy.  This process is complex and involves over 20 different proteins, most of which are highly conserved acro...  Read full blog post.

The role of Parkin and autophagy in retinal pigment epithelial cell (RPE) degradation
The root of Parkinson’s disease (PD) points to a poorly regulated electron transport chain leading to mitochondrial damage, where many proteins need to work cohesively to ensure proper function.  The two key players of this pathway are PINK1, ...  Read full blog post.

The identification of dopaminergic neurons using Tyrosine Hydroxylase in Parkinson's research and LRRK2
Tyrosine hydroxylase (TH) is a crucial enzyme involved in the biosynthesis of dopamine, norepinephrine and epinephrine in the brain.  Specifically, TH catalyzes the conversion of l-tyrosine to l-dihydroxyphenylalanine (l-dopa).  The importance of t...  Read full blog post.

Parkin - Role in Mitochondrial Quality Control and Parkinson's Disease
Parkin/PARK2 is a cytosolic enzyme which gets recruited to cellular mitochondria damaged through depolarization, ROS or unfolded proteins accumulation, and exert protective effects by inducing mitophagy (mitochondrial autophagy). Parkin induces mit...  Read full blog post.

PINK1 - performing mitochondrial quality control and protecting against Parkinson’s disease
PTEN-induced putative kinase 1 (PINK1) is a serine/threonine kinase with important functions in mitochondrial quality control. Together with the Parkin protein, PINK1 is able to regulate the selective degradation of damaged mitochondria through aut...  Read full blog post.

PINK1: All work and no fun
The protein PINK1 is a mitochondrial-located serine/threonine kinase (PTK) that maintains organelle function and integrity. It not only protects organelles from cellular stress, but it also uses the selective auto-phagocytosis process for cleaning and...  Read full blog post.

PINK1 and its role in Parkinson's disease
PINK1 (PTEN induced putative kinase 1) is a mitochondrial serine/threonine kinase which maintains mitochondrial function/integrity, provides protection against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondr...  Read full blog post.

Showing 1-10 of 14 blog posts - Show all blog posts.
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Recent Reviews

3.9
5
4
4
6
3
2
2
0
1
1

Verified Customer
11/08/2021
Application: WB
Species: Human

Verified Customer
01/09/2021
Application: WB
Species: Mouse

Muhammad Yousuf
03/23/2019
Application: WB
Species: Mouse

Bioinformatics

Gene Symbol PINK1
Entrez
Uniprot