Anti-PDK4 antibody [EPR19727-245]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PDK4 antibody [EPR19727-245] (AB214938)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed C2C12 (Mouse myoblast cell line) cells labeling PDK4 with ab214938 at 1/60 dilution (red) compared withRabbit 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 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-PDK4 antibody [EPR19727-245] (AB214938)

PDK4 was immunoprecipitated from 0.35 mg of mouse heart lysate with ab214938 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab214938 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1 : Mouse heart lysate 10μg (Input).

Lane 2 : ab214938 IP in mouse heart lysate.

Lane 3 : Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab214938 in mouse heart lysate.

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

Exposure time : 0.5 second.

All lanes:

Immunoprecipitation - Anti-PDK4 antibody [EPR19727-245] (ab214938)

Predicted band size: 46 kDa,48 kDa

false

• WB

Supplier Data

Western blot - Anti-PDK4 antibody [EPR19727-245] (AB214938)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 5 seconds; Lane 2-4 : 3 minutes; Lane 5 : 1 minutes; Lane 6 : 10 seconds.

All lanes:

Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/1000 dilution

Lane 1:

Mouse skeletal muscle lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Rat muscle lysate at 10 µg

Lane 4:

Rat heart lysate at 10 µg

Lane 5:

C2C12 (Mouse myoblast cell line) whole cell lysate at 10 µg

Lane 6:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 47 kDa

false

• WB

Supplier Data

Western blot - Anti-PDK4 antibody [EPR19727-245] (AB214938)

Blocking/Dilution buffer : 5% NFDM/TBST.

Mouse PDK4, PDK1 and PDK2 recombinant proteins contain aa8-412, aa1-434 and aa1-407 respectively with a His-Tag®.

All lanes:

Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/10000 dilution

Lane 1:

Mouse PDK4 recombinant protein at 0.01 µg

Lane 2:

Mouse PDK1 recombinant protein at 0.01 µg

Lane 3:

Mouse PDK2 recombinant protein at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 47 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-PDK4 antibody [EPR19727-245] (AB214938)

Blocking/Dilution buffer : 5% NFDM/TBST.

The expression level of PDK4 is increased under the fasting state (PMID : 11723055 and PMID : 16606348).

Lanes 1 - 2:

Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/10000 dilution

Lanes 3 - 4:

Western blot - Anti-PDK4 antibody [EPR19727-245] (ab214938) at 1/1000 dilution

Lane 1:

Heart from nonfasting mouse at 10 µg

Lane 2:

Heart from mouse fasting for 72 hours at 10 µg

Lane 3:

Gastrocnemius muscle from nonfasting mouse at 10 µg

Lane 4:

Gastrocnemius muscle from mouse fasting for 72 hours at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 46 kDa

Observed band size: 47 kDa

false

Exposure time: 5s

• WB

CiteAb

Western blot - Anti-PDK4 antibody [EPR19727-245] (AB214938)

PDK4 western blot using anti-PDK4 antibody [EPR19727-245] ab214938. Publication image and figure legend from Gao, K., Zhang, J., et al., 2020, Chin Med, PubMed 32158496.

ab214938 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab214938 please see the product overview.

Effects of QSG on the uncoupling of glycolysis from glucose oxidation in HF rats after AMI. a Lactate level in serum and myocardial tissue. Lactate level in serum and myocardial tissue was significantly increased following HF and downregulated with QSG treatment. n = 8 per group. Values are mean ± SE. Asterisks indicates significant differences. *p < 0.05, **p < 0.01. b Myocardial protein expressional levels of LDHA and PDK4 in the three groups. LDHA involved in regulating glycolysis was observably elevated in the model group. LDHA level was significantly down-regulated by treatment of QSG compared with the model group. PDK4, decreasing glucose oxidation by an inhibitory phosphorylation of the pyruvate dehydrogenase complex, was up-regulated in the model group and down-regulated in the QSG group. n = 4 per group. Values are mean ± SE. Asterisks indicates significant differences. *p < 0.05, **p < 0.01

false