Antibodies, Proteins, Kits and Reagents for Life Science

278 083 ₸ Product size

100 µl 278 083 ₸

Order now and get it on Tuesday March 02, 2021

Anti-PINK1 antibody [EPR20730] (ab216144)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20730] to PINK1
Suitable for: WB, ICC/IF, IP
Reacts with: Human

Product name

Anti-PINK1 antibody [EPR20730]
See all PINK1 primary antibodies
Description

Rabbit monoclonal [EPR20730] to PINK1
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa cells (+/- treatment with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate; human PINK1 recombinant protein (aa156-507) . ICC/IF: HeLa cells treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. IP: HeLA cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate.
General notes

Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20730
Isotype

IgG
Research areas

Western blot - Anti-PINK1 antibody [EPR20730] (ab216144)

Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution + Human PINK1 recombinant protein (aa156-507), 10 ng

Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution (Goat Anti-Rabbit IgG H&L (HRP))

Developed using the ECL technique.

Predicted band size: 63 kDa
Observed band size: 38 kDa
why is the actual band size different from the predicted?

Exposure time: 1 second

Dilution/blocking buffer: 5% NFDM/TBST
Immunocytochemistry/ Immunofluorescence - Anti-PINK1 antibody [EPR20730] (ab216144)

Immunofluorescent analysis of 4 % paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma)(+/- treatment with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) cells labeling PINK1 with ab216144 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells treated with 10μM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

The negative controls are as follows:
-ve control: PBS, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.
Western blot - Anti-PINK1 antibody [EPR20730] (ab216144)

All lanes : Anti-PINK1 antibody [EPR20730] (ab216144) at 1/1000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa cells (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP, ab141229) for 24 hours) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Developed using the ECL technique.

Predicted band size: 63 kDa
Observed band size: 62 kDa why is the actual band size different from the predicted?

Exposure time: 5 seconds

Blocking and dilution buffer: 5% NFDM/TBST

PINK1 can be induced by CCCP treatment (PMID: 24184327).
Immunoprecipitation - Anti-PINK1 antibody [EPR20730] (ab216144)

PINK1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) (treated with 10uM carbonyl cyanide 3-chlorophenylhydrazone (CCCP. ab141229) for 24 hours) whole cell lysate with ab216144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216144 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.

Lane 1: HeLa (CCCP-treated, ab141229) lysate 10 μg (Input).
Lane 2: ab216144 IP in HeLa (CCCP-treated, ab141229) lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216144 in HeLa (CCCP-treated, ab141229) whole cell lysate.

Blocking and dilution buffer: 5% NFDM/TBST.
Anti-PINK1 antibody [EPR20730] (ab216144)

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