Anti-TGF beta Receptor I antibody [EPR20923-13]

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (AB235578)

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.

All lanes:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (ab235578) 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

• IP

Supplier Data

Immunoprecipitation - Anti-TGF beta Receptor I antibody [EPR20923-13] (AB235578)

TGF beta Receptor I was immunoprecipitated from 0.35 mg MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate 10 μg 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.

All lanes:

Immunoprecipitation - Anti-TGF beta Receptor I antibody [EPR20923-13] (ab235578)

Predicted band size: 55 kDa

Observed band size: 55 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (AB235578)

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] (ab235578) 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] (AB235578)

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

Exposure time : 3 minutes

All lanes:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] (ab235578) 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] (AB235578)

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] (ab235578) 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] (AB235578)

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] (ab235578) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-TGF beta Receptor I antibody [EPR20923-13] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-i-antibody-epr20923-13-bsa-and-azide-free-ab288303'>ab288303</a>) 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