Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (AB288303)

TGF beta Receptor I was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate with ab235578 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab235578 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab235578 IP in MCF7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab235578 in MCF7 whole cell lysate

The bands beneath the target band is caused by degradation as demonstrated by WB data.

This data was developed using ab235578, the same antibody clone in a different buffer formulation.

All lanes:

Immunoprecipitation - Anti-TGF beta Receptor I antibody [EPR20923-13] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-ab235578'>ab235578</a>)

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (AB288303)

This data was developed using ab235578, the same antibody clone in a different buffer formulation.

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

Lysate were made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-ab235578'>ab235578</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

2.4G2 (rat B cell lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 4:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 55 kDa

Observed band size: 37 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (AB288303)

This data was developed using ab235578, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration :

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-ab235578'>ab235578</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

MCF7 transfected with 25nM siRNA specifically targeti TGF receptor I whole cell lysate at 20 µg

Lane 3:

MCF7 transfected with 100nM siRNA specifically targeti TGF receptor I whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (AB288303)

This data was developed using ab235578, the same antibody clone in a different buffer formulation.

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

Lane1 Lysate was made freshly and used in WB test immediately to minimize protein degradation.

Exposure time : 81 seconds

All lanes:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-ab235578'>ab235578</a>) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell fresh lysate at 20 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/50000 dilution

Predicted band size: 55 kDa

Observed band size: 37 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (AB288303)

False colour image of Western blot : Anti-TGF beta Receptor I antibody [EPR20923-13] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab235578 was shown to bind specifically to TGF beta Receptor I. A band was observed at 40/55 kDa in wild-type A549 cell lysates with no signal observed at this size in TGFBR1 knockout cell line ab277894 (knockout cell lysate ab283082). To generate this image, wild-type and TGFBR1 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using ab235578, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-ab235578'>ab235578</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

Western blot - Human TGFBR1 knockout A549 cell lysate (ab283082) at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Predicted band size: 55 kDa

Observed band size: 40 kDa,55 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (AB288303)

This data was developed using ab235578, the same antibody clone in a different buffer formulation.

Western blot : Anti-TGF beta Receptor I antibody [EPR20923-13] ab235578 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 42 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in TGFBR1 knockout HCT 116 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

Lanes 1 - 3:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-ab235578'>ab235578</a>) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (ab288303) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human TGFBR1 knockout HCT116 cell line (ab289125) at 20 µg

Lane 3:

A-431 UNBOILED at 20 µg

Secondary

All lanes:

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

Predicted band size: 56 kDa

Observed band size: 42 kDa

false