Anti-Cdk6 antibody [98D] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-Cdk6 antibody [98D] - BSA and Azide free (AB252796)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

Negative control : PANC-1, Saos-2. (PMID : 8114739).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241554).

All lanes:

Western blot - Anti-Cdk6 antibody [98D] (<a href='/en-us/products/primary-antibodies/cdk6-antibody-98d-ab241554'>ab241554</a>) at 0.518 µg/mL

Lane 1:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 20 µg

Lane 3:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

Saos-2 (human osteosarcoma epithelial), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 37 kDa

Observed band size: 37 kDa

false

• WB

Lab

Western blot - Anti-Cdk6 antibody [98D] - BSA and Azide free (AB252796)

This data was developed using the same antibody clone in a different buffer formulation (ab241554).

Lanes 1 - 2 : Merged signal (red and green). Green - ab241554 observed at 38 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab241554 was shown to react with Cdk6 in HeLa wild-type cells in Western blot with loss of signal observed in CDK6 knockout cell line ab266059 (CDK6 knockout cell lysate ab257088). Wild-type HeLa and CDK6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab241554 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Cdk6 antibody [98D] (<a href='/en-us/products/primary-antibodies/cdk6-antibody-98d-ab241554'>ab241554</a>) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CDK6 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CDK6 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cdk6-knockout-hela-cell-line-ab266059'>ab266059</a>)

Predicted band size: 37 kDa

Observed band size: 38 kDa

false

• WB

Supplier Data

Western blot - Anti-Cdk6 antibody [98D] - BSA and Azide free (AB252796)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241554).

All lanes:

Western blot - Anti-Cdk6 antibody [98D] (<a href='/en-us/products/primary-antibodies/cdk6-antibody-98d-ab241554'>ab241554</a>) at 0.518 µg/mL

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 2:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Predicted band size: 37 kDa

Observed band size: 37 kDa

false