Anti-PDK2 antibody [EPR1948Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDK2 antibody [EPR1948Y] (AB68164)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cardiac muscle tissue sections labeling PDK2 with Purified ab68164 at 1/1000 dilution (0.12 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PDK2 antibody [EPR1948Y] (AB68164)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human bladder carcinoma tissue sections labeling PDK2 with Purified ab68164 at 1/1000 dilution (0.12 µg/mL). Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

Lab

Immunoprecipitation - Anti-PDK2 antibody [EPR1948Y] (AB68164)

Purified ab68164 at 1/20 dilution (0.6µg) immunoprecipitating PDK2 in Rat heart lysate.
Lane 1 (input) : Rat heart lysate 10µg
Lane 2 (+) : ab68164 + Rat heart lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab68164 in Rat heart lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 48 kDa

All lanes:

Immunoprecipitation - Anti-PDK2 antibody [EPR1948Y] (ab68164)

Predicted band size: 106 kDa,46 kDa

false

• WB

Lab

Western blot - Anti-PDK2 antibody [EPR1948Y] (AB68164)

This data was developed using ab227847, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal[EPR1948Y] to PDK2 ab227847 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in PDK2 knockout U-87 MG cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-PDK2 antibody [EPR1948Y] - BSA and Azide free (<a href='/ko/products/primary-antibodies/pdk2-antibody-epr1948y-bsa-and-azide-free-ab227847'>ab227847</a>) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

PDK2 knockout U-87 MG at 20 µg

Lane 3:

SH-SY5Y at 20 µg

Lane 4:

HL-60 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 45 kDa

Observed band size: 46 kDa,36 kDa

false

• WB

Unknown

Western blot - Anti-PDK2 antibody [EPR1948Y] (AB68164)

All lanes:

Western blot - Anti-PDK2 antibody [EPR1948Y] (ab68164) at 1/200 dilution

All lanes:

Human heart lysate

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 46 kDa

false

• WB

Unknown

Western blot - Anti-PDK2 antibody [EPR1948Y] (AB68164)

All lanes:

Western blot - Anti-PDK2 antibody [EPR1948Y] (ab68164) at 1/1000 dilution

Lane 1:

Mouse heart lysate at 20 µg

Lane 2:

Rat heart lysate at 20 µg

Lane 3:

Rat skeletal muscle lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 46 kDa

false