Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free (AB309097)

This data was developed using ab309096, the same antibody clone in a different buffer formulation. METTL14 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab309096 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab309096 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate Lane 2 : ab309096 IP in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab309096 in Raji whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-METTL14 antibody [EPR27234-62] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr27234-62-ab309096'>ab309096</a>) at 1/30 dilution

All lanes:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 60 kDa

false

Exposure time: 41s

• WB

Supplier Data

Western blot - Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free (AB309097)

This data was developed using ab309096, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-METTL14 antibody [EPR27234-62] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr27234-62-ab309096'>ab309096</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human colon tissue lysate at 20 µg

Lane 3:

Human colon cancer tissue 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 1/20000 dilution

Observed band size: 60 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free (AB309097)

This data was developed using ab309096, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the bands between 100 kDa and 300 kDa are unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-METTL14 antibody [EPR27234-62] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr27234-62-ab309096'>ab309096</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeti METTL14 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 1/20000 dilution

Observed band size: 60 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free (AB309097)

This data was developed using ab309096, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. The identity of the bands between 150 kDa and 300 kDa are unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 48 seconds

All lanes:

Western blot - Anti-METTL14 antibody [EPR27234-62] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr27234-62-ab309096'>ab309096</a>) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) 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 1/20000 dilution

Observed band size: 60 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free (AB309097)

This data was developed using ab309096, the same antibody clone in a different buffer formulation.

Exposure time : ab300104 : 3 seconds; ab309096 : 40 seconds.

ab300104 is highly sensitive but unfortunately detects a nonspecific band near the METTL14 target band.

Lanes 1 - 7:

Western blot - Anti-METTL14 antibody [EPR22666-22] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr22666-22-ab300104'>ab300104</a>) at 1/1000 dilution

Lanes 8 - 14:

Western blot - Anti-METTL14 antibody [EPR27234-62] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr27234-62-ab309096'>ab309096</a>) at 1/1000 dilution

Lanes 1 and 8:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lanes 2 and 9:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 3 and 10:

293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lanes 4 and 11:

Human testis tissue lysate at 20 µg

Lanes 5 and 12:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lanes 6 and 13:

Mouse testis at 20 µg

Lanes 7 and 14:

Mouse lung at 20 µg

Secondary

All lanes:

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

Observed band size: 60 kDa

false

• WB

Supplier Data

Western blot - Anti-METTL14 antibody [EPR27234-62] - BSA and Azide free (AB309097)

This data was developed using ab309096, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST The identity of the bands between 150 kDa and 300 kDa are unknown. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : Lane 1 : 48 seconds, lanes 2-3 : 180 seconds.

All lanes:

Western blot - Anti-METTL14 antibody [EPR27234-62] (<a href='/en-us/products/primary-antibodies/mettl14-antibody-epr27234-62-ab309096'>ab309096</a>) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Rat testis tissue lysate at 20 µg

Lane 3:

Mouse lu tissue 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 1/20000 dilution

Observed band size: 60 kDa

false