Anti-CNOT6 antibody [EPR22022]

• IP

Supplier Data

Immunoprecipitation - Anti-CNOT6 antibody [EPR22022] (AB221151)

CNOT6 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab221151 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221151 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

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

Lane 2 : ab221151 IP in HeLa whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221151 in HeLa whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-CNOT6 antibody [EPR22022] (ab221151)

Predicted band size: 36 kDa,63 kDa

Observed band size: 38 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-CNOT6 antibody [EPR22022] (AB221151)

CNOT6 was immunoprecipitated from 0.35 mg of Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate with ab221151 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221151 at 1/500 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1000 dilution.

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

Lane 2 : ab221151 IP in Jurkat whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221151 in Jurkat whole cell lysate.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-CNOT6 antibody [EPR22022] (ab221151)

Predicted band size: 63 kDa

false

• WB

Supplier Data

Western blot - Anti-CNOT6 antibody [EPR22022] (AB221151)

Blocking/Dilution buffer : 5% NFDM/TBST.

The reactivity of the antibody with the target protein in the HeLa lysate was blocked by the inclusion of the peptide antigen.

All lanes:

Western blot - Anti-CNOT6 antibody [EPR22022] (ab221151) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

HeLa whole cell lysate at 20 µg with CNOT6 Immunogen peptide

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: 63 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-CNOT6 antibody [EPR22022] (AB221151)

Western blot : Anti-CNOT6 antibody [EPR22022] ab221151 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 63 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in CNOT6 knockout MCF7 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-CNOT6 antibody [EPR22022] (ab221151) at 1/1000 dilution

Lane 1:

Wild-type MCF7 at 20 µg

Lane 2:

Western blot - Human CNOT6 knockout MCF7 cell line (ab289386) at 20 µg

Lane 3:

HL-60 at 20 µg

Lane 4:

PC-3 at 20 µg

Secondary

All lanes:

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

Predicted band size: 63 kDa

Observed band size: 63 kDa,37 kDa

false

• WB

Lab

Western blot - Anti-CNOT6 antibody [EPR22022] (AB221151)

The expression profile is consistent with what has been described in the literature (PMID : 21741365).

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

All lanes:

Western blot - Anti-CNOT6 antibody [EPR22022] (ab221151) at 1/1000 dilution

Lane 1:

Human testis lysate at 20 µg

Lane 2:

Human ovary lysate at 20 µg

Lane 3:

Human heart lysate at 20 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 63 kDa

Observed band size: 60 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CNOT6 antibody [EPR22022] (AB221151)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CNOT6 antibody [EPR22022] (ab221151) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Human ovary cancer lysate at 20 µg

Secondary

Lanes 1 - 3:

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

Lane 4:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 63 kDa

false

Exposure time: 3min

• WB

Lab

Western blot - Anti-CNOT6 antibody [EPR22022] (AB221151)

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

All lanes:

Western blot - Anti-CNOT6 antibody [EPR22022] (ab221151) at 1/1000 dilution

Lane 1:

His-tagged human CNOT6 recombinant protein(aa1-210) at 0.002 µg

Lane 2:

His-tagged human CNOT6L recombinant protein(aa1-210) at 0.002 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: 63 kDa

false

Exposure time: 3s