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 [EPR19573] (ab207450)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19573] to PDK1
Suitable for: WB, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-PDK1 antibody [EPR19573]
See all PDK1 primary antibodies
Description

Rabbit monoclonal [EPR19573] to PDK1
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Human

WB
Mouse

Rat

Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa, Hap1, HEK-293, Jurkat, HepG2, NIH/3T3, PC-12 and LNCaP whole cell lysates; human heart and kidney lysates; mouse brain, heart, kidney and spleen lysates; rat kidney and spleen lysates; HEK-293 mitochondria fraction lysate. IP: HepG2 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.

Application	Species
IP	Human
WB	Mouse Rat Human

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, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19573
Isotype

IgG
Research areas

Western blot - Anti-PDK1 antibody [EPR19573] (ab207450)

All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PDK1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 49 kDa
Observed band size: 45 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab207450 observed at 45 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab207450 was shown to react with PDK1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264791 (knockout cell lysate ab257164) was used. Wild-type HeLa and PDK1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab207450 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-PDK1 antibody [EPR19573] (ab207450)

All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution

Lane 1 : Wild-type HAP1 cell lysate
Lane 2 : PDK1 knockout HAP1 cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : LnCaP cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 49 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab207450 observed at 45 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab207450 was shown to specifically react with PDK1 in wild-type HAP1 cells. No band was observed when PDK1 knockout samples were examined. Wild-type and PDK1 knockout samples were subjected to SDS-PAGE. Ab207450 and ab8245 (loading control to GAPDH) were diluted at 1/2000 and 1/10,000 dilution respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Immunoprecipitation - Anti-PDK1 antibody [EPR19573] (ab207450)

PDK1 was immunoprecipitated from 1mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab207450 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab207450 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: HepG2 whole cell lysate 10µg (Input).

Lane 2: ab207450 IP in HepG2 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab207450 in HepG2 whole cell lysate.

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

Exposure time: 1 second.
Western blot - Anti-PDK1 antibody [EPR19573] (ab207450)

All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 5 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 6 : LNCaP (Human prostate cancer cell line) whole cell 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 1-5: 15 seconds; Lane 6: 8 seconds.
Western blot - Anti-PDK1 antibody [EPR19573] (ab207450)

All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/2000 dilution

Lane 1 : Human heart lysate
Lane 2 : Human kidney lysate
Lane 3 : Mouse brain lysate
Lane 4 : Mouse heart lysate
Lane 5 : Mouse kidney lysate
Lane 6 : Mouse spleen lysate
Lane 7 : Rat kidney lysate
Lane 8 : Rat spleen lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Lanes 3-8 : 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 1/3/4: 8 seconds; Lane 2: 30 seconds; Lane 5/6: 3 minutes; Lane 7/8: 15 seconds.
Western blot - Anti-PDK1 antibody [EPR19573] (ab207450)

All lanes : Anti-PDK1 antibody [EPR19573] (ab207450) at 1/5000 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) mitochondria fraction lysate
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) cytosol fraction 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?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Anti-PDK1 antibody [EPR19573] (ab207450)

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