Anti-PKN1 antibody [EPR18808] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-PKN1 antibody [EPR18808] - BSA and Azide free (AB251202)

This data was developed using ab195264, the same antibody clone in a different buffer formulation.

PKN1 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate with ab195264 at 1/40 dilution.

Western blot was performed from the immunoprecipitate using ab195264 at 1/1000 dilution.

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

Lane 1 : Jurkat whole cell lysate, 10μg (Input).
Lane 2 : ab195264 IP in Jurkat whole cell lysate.
Lane 3 : Rabbit IgG,monoclonal-Isotype Control (ab172730) instead of ab195264 in Jurkat whole cell lysate.

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-PKN1 antibody [EPR18808] (<a href='/en-us/products/primary-antibodies/pkn1-antibody-epr18808-ab195264'>ab195264</a>)

Predicted band size: 104 kDa

false

• WB

Lab

Western blot - Anti-PKN1 antibody [EPR18808] - BSA and Azide free (AB251202)

This data was developed using ab195264, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (40 μg)

Lane 2 : PKN1 knockout HAP1 cell lysate (40 μg)

Lane 3 : MCF7 cell lysate (20 μg)

Lane 4 : K562 cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab195264 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab195264 was shown to specifically react with PKN1 when PKN1 knockout samples were used. Wild-type and PKN1 knockout samples were subjected to SDS-PAGE. ab195264 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PKN1 antibody [EPR18808] (<a href='/en-us/products/primary-antibodies/pkn1-antibody-epr18808-ab195264'>ab195264</a>)

Predicted band size: 104 kDa

false

• WB

Supplier Data

Western blot - Anti-PKN1 antibody [EPR18808] - BSA and Azide free (AB251202)

This data was developed using ab195264, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKN1 antibody [EPR18808] (<a href='/en-us/products/primary-antibodies/pkn1-antibody-epr18808-ab195264'>ab195264</a>) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg

Lane 3:

MDA-MB-231 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 20 µg

Lane 5:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 6:

MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 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: 104 kDa

Observed band size: 120 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PKN1 antibody [EPR18808] - BSA and Azide free (AB251202)

This data was developed using ab195264, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-PKN1 antibody [EPR18808] (<a href='/en-us/products/primary-antibodies/pkn1-antibody-epr18808-ab195264'>ab195264</a>) at 1/1000 dilution

Lane 1:

Rat brain lysate at 10 µg

Lane 2:

Rat spleen 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: 104 kDa

Observed band size: 120 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-PKN1 antibody [EPR18808] - BSA and Azide free (AB251202)

This data was developed using ab195264, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PKN1 antibody [EPR18808] (<a href='/en-us/products/primary-antibodies/pkn1-antibody-epr18808-ab195264'>ab195264</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast 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: 104 kDa

Observed band size: 120 kDa

false

Exposure time: 3min