Antibodies, Proteins, Kits and Reagents for Life Science

324 988 ₸ Product size

100 µl 324 988 ₸ 10 µl 130 665 ₸

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Anti-PDK1 antibody [EPR19571] (ab202468)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19571] to PDK1
Suitable for: WB, ICC/IF, Flow Cyt, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-PDK1 antibody [EPR19571]
See all PDK1 primary antibodies
Description

Rabbit monoclonal [EPR19571] to PDK1
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: L-929, NIH/3T3, F9, bEnd.3, C6 and PC-12 whole cell lysates; Mouse brain, kidney, spleen and heart lysates; Rat brain, kidney and heart lysates. ICC/IF: NIH/3T3 and RAW 264.7 cells. Flow Cyt: NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
General notes
This product was previously labelled as Mitochondrial Pyruvate dehydrogenase kinase 1

This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
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: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19571
Isotype

IgG
Research areas

Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)

Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: PDK1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: Jurkat whole cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab202468 observed at 45 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab202468 was shown to specifically react with PDK1 in wild-type cells along with additional cross reactive bands. Signal was lost in PDK1 knockout cells. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. ab202468 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/2000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PDK1 with ab202468 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on NIH/3T3 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab202468 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Flow Cytometry - Anti-PDK1 antibody [EPR19571] (ab202468)

Flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling PDK1 with ab202468 at 1/600 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/500 dilution was used as the secondary antibody.

Cells were permeabilised by 90% methanol-1XPBS, -20ºC, 30min.
Immunoprecipitation - Anti-PDK1 antibody [EPR19571] (ab202468)

PDK1 was immunoprecipitated from 1mg of NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate with ab202468 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab202468 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: NIH/3T3 whole cell lysate, 10µg (Input).

Lane 2: ab202468 IP in NIH/3T3 whole cell lysate.

Lane 3: Rabbit IgG,monoclonal -[EPR25A] - Isotype Control (ab172730) instead of ab202468 in NIH/3T3 whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 1 second.
Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)

All lanes : Anti-PDK1 antibody [EPR19571] (ab202468) at 1/2000 dilution

Lane 1 : L-929 (Mouse connective tissue fibroblast cell line) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 3 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate
Lane 4 : bEnd.3 (Mouse brain endothelioma cell line) whole cell lysate
Lane 5 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 6 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 49 kDa
Observed band size: 47 kDa
why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 1 minute; Lane 3-4: 10 seconds; Lane 5/6: 8 seconds.
Immunocytochemistry/ Immunofluorescence - Anti-PDK1 antibody [EPR19571] (ab202468)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) cells labeling PDK1 with ab202468 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on RAW 264.7 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab202468 at 1/1000 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2: ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.
Western blot - Anti-PDK1 antibody [EPR19571] (ab202468)

All lanes : Anti-PDK1 antibody [EPR19571] (ab202468) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Mouse heart lysate
Lane 5 : Rat brain lysate
Lane 6 : Rat kidney lysate
Lane 7 : Rat heart lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 49 kDa
Observed band size: 47 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1/2/3/5/6: 8 seconds; Lane 4: 3 seconds; Lane 7: 1 second.
Anti-PDK1 antibody [EPR19571] (ab202468)

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