Overview

  • Product name
  • Description
    Rabbit polyclonal to PINK1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Cynomolgus monkey
  • Immunogen

    Synthetic peptide:

    LVRALLQREA SKRPSARVAA N

    , corresponding to amino acids 484-504 of Human PINK1.

  • Positive control
    • Human, mouse, rat liver.

Properties

Applications

Our Abpromise guarantee covers the use of ab23707 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/200 - 1/1000. Detects a band of approximately 66 kDa.Can be blocked with Human PINK1 peptide (ab30903). Detects a band of approximately 66 kDa. Can be blocked with PINK1 peptide (484-504) (ab30903).
IHC-P Use a concentration of 4 µg/ml.
ICC/IF Use a concentration of 1 - 5 µg/ml.

Target

  • Function
    Protects against mitochondrial dysfunction during cellular stress, potentially by phosphorylating mitochondrial proteins. Involved in the clearance of damaged mitochondria via selective autophagy (mitophagy). It is necessary for PARK2 recruitement to dysfunctional mitochondria to initiate their degradation.
  • Tissue specificity
    Highly expressed in heart, skeletal muscle and testis, and at lower levels in brain, placenta, liver, kidney, pancreas, prostate, ovary and small intestine. Present in the embryonic testis from an early stage of development.
  • Involvement in disease
    Defects in PINK1 are the cause of Parkinson disease type 6 (PARK6) [MIM:605909]. A neurodegenerative disorder characterized by parkinsonian signs such as rigidity, resting tremor and bradykinesia. A subset of patients manifest additional symptoms including hyperreflexia, autonomic instability, dementia and psychiatric disturbances. Symptoms show diurnal fluctuation and can improve after sleep.
  • Sequence similarities
    Belongs to the protein kinase superfamily. Ser/Thr protein kinase family.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Autophosphorylated.
  • Cellular localization
    Mitochondrion outer membrane. Cytoplasm > cytosol.
  • Information by UniProt
  • Database links
  • Alternative names
    • BRPK antibody
    • FLJ27236 antibody
    • mitochondrial antibody
    • PARK 6 antibody
    • PARK6 antibody
    • Phosphatase and Tensin Homolog antibody
    • PINK 1 antibody
    • PINK1 antibody
    • PINK1_HUMAN antibody
    • Protein kinase BRPK antibody
    • PTEN induced putative kinase 1 antibody
    • PTEN induced putative kinase protein 1 antibody
    • PTEN-induced putative kinase protein 1 antibody
    • Serine/threonine kinase PINK1 mitochondrial antibody
    • Serine/threonine protein kinase PINK1 mitochondrial antibody
    • Serine/threonine-protein kinase PINK1 antibody
    see all

Images

  • All lanes : Anti-PINK1 antibody (ab23707) at 1/1000 dilution

    Lane 1 : Human liver whole cell lysate
    Lane 2 : Human hepatocytes whole cell lysate

    Lysates/proteins at 25 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP conjugate at 1/1000 dilution

    Developed using the ECL technique.

    Observed band size: 66 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 50 kDa (possible non-specific binding)


    Exposure time: 20 seconds


    Blocking: 5% milk for 1 hour at 23°C.

    See Abreview

  • Anti-PINK1 antibody (ab23707) at 4 µg/ml + Murine liver 100,000 x g pellet at 30 µg

    Observed band size: 66 kDa why is the actual band size different from the predicted?
    Additional bands at: 33 kDa (possible cleavage fragment)

  • Anti-PINK1 antibody (ab23707) at 1/1000 dilution + Whole cell lysate prepared from Jurkat cells at 100000 cells

    Secondary
    Donkey anti-rabbit IgG conjugated to HRP at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 66 kDa why is the actual band size different from the predicted?
    Additional bands at: 100 kDa, 35 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 10 seconds


    Samples blocked with 5% milk for 1 hour at 25°C.

    See Abreview

  • ICC/IF image of ab23707 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab23707, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References

This product has been referenced in:
  • Baumgartner C  et al. An ERK-Dependent Feedback Mechanism Prevents Hematopoietic Stem Cell Exhaustion. Cell Stem Cell 22:879-892.e6 (2018). Read more (PubMed: 29804890) »
  • Deyu H  et al. Protective mechanisms involving enhanced mitochondrial functions and mitophagy against T-2 toxin-induced toxicities in GH3 cells. Toxicol Lett 295:41-53 (2018). Read more (PubMed: 29870751) »
See all 35 Publications for this product

Customer reviews and Q&As

1-10 of 28 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (head and neck cancer cell)
Gel Running Conditions
Reduced Denaturing (8% SDS page gel)
Loading amount
30 µg
Treatment
10uM CCCP for 24h
Specification
head and neck cancer cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Sep 19 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Colon adenocarcinoma)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 8.5 buffer
Permeabilization
No
Specification
Colon adenocarcinoma
Fixative
Formaldehyde

Mr. Allan Wang

Verified customer

Submitted Mar 20 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (human hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
human hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 26 2017

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - Triton x-100, 0.01%
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jun 19 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Permeabilization
Yes - Triton x-100, 0.01%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 30 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - nuclear (Cardiomyocytes)
Specification
Cardiomyocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 02 2015

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Sample
Mouse Cell (Cardiomyocytes)
Specification
Cardiomyocytes
Permeabilization
Yes - Triton x-100, 0.01%
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Feb 20 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 04 2014

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 04 2014

Answer



I have contacted the laboratories who have provided the IHC-P testing protocol which I hope will be helpful. Please note this would be a guideline only and may require individual optimization. I have copied it below.




Immunostaining Procedure:

Deparaffinize sections with Xylene or Xylene substitute 3 times, 5 minutes each.
Rehydrate sections with 100% ethanol 2 times, 5 minutes each, followed by 95%, 90%, 80%, 70%, 50% ethanol, 5 minutes each.
Rinse sections in distilled water for 5 minutes.
Block endogenous peroxidase activity with 0.3% H2O2 in water (use methanol instead of water in case of strong endogenous peroxidase activity) for 15 minutes.
Wash sections in TBS containing 1% tween 20 (TBST), pH 7.4, 3 times, 5 minutes each.
Incubate sections with 5% normal serum (same species as the host of secondary antibody) for 30 minutes.
Rinse sections with TBST, pH 7.4, 3 times, 5 minutes each.
Incubate sections with 1:100 Antibody (or recommended starting dilution. Optimal dilution to be determined by end user) overnight at 40C.
Wash sections in TBST, pH 7.4, 3 times, 5 minutes each.
Incubate sections for 30 minutes with biotinylated secondary antibody, using a dilution as recommended by provider.
Wash sections in TBST, pH 7.4, 3 times, 5 minutes each.
Incubate sections for 30 minutes with ABC reagent, using a dilution as recommended by provider.
Wash sections in TBST, pH 7.4, 3 times, 5 minutes each.
Incubate sections in peroxidase substrate solution. Check staining under a microscope frequently. When desired staining intensity is achieved, rinse sections with distilled water thoroughly.
Counter stain sections if desired. Rinse sections thoroughly after counter stain.





Fixation:

Fresh tissues and put them into 4% paraformaldehyde in PBS overnight at 4C.

Read More

1-10 of 28 Abreviews or Q&A

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