Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab259360, the same antibody clone in a different buffer formulation.

TGF beta Receptor II was immunoprecipitated from 0.35 mg HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate 10 μg with ab259360 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259360 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab259360 IP in HT-1080 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259360 in HT-1080 whole cell lysate

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

Exposure time : 76 seconds.

Bands below 50 kDa are caused by degradation as demonstrated by WB data.

All lanes:

Immunoprecipitation - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>)

Predicted band size: 65 kDa

Observed band size: 70-80 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab259360, the same antibody clone in a different buffer formulation.

TGF beta Receptor II was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg with ab259360 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab259360 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg

Lane 2 : ab259360 IP in NIH/3T3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab259360 in NIH/3T3 whole cell lysate

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

Exposure time : 50 seconds.

Bands below 50 kDa are caused by degradation as demonstrated by WB data.

All lanes:

Immunoprecipitation - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>)

Predicted band size: 65 kDa

Observed band size: 70-80 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab259360, the same antibody clone in a different buffer formulation.

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

Negative control : MCF7, PC-12 (PMID : 1310899).

Lysates used in this blot have experienced freeze-thaw cycles. Bands below 50 kDa are caused by degradation.

Exposure time : Lanes 1-4 : 81 seconds; Lane 5 : 3 minutes.

All lanes:

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>) at 1/1000 dilution

Lane 1:

HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

L6 (rat skeletal muscle myoblast) whole cell lysate at 20 µg

Lane 4:

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

Lane 5:

Rat lung 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 1/100000 dilution

Predicted band size: 65 kDa

Observed band size: 70-80 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab259360, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST. The molecular weight observed is consistent with what has been described in the literature (PMID : 23468914).

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

Exposure time : Lanes 1-2 : 81 seconds;Lane 3 : 3 minutes.

All lanes:

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>) at 1/1000 dilution

Lane 1:

HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell) 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 1/100000 dilution

Predicted band size: 65 kDa

Observed band size: 70-80 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using the same antibody clone in a different buffer formulation (ab259360).

Western blot : Anti-TGFBR2 antibody [EPR24349-124] (ab259360) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab259360 was shown to bind specifically to TGFBR2. A band was observed at 65 kDa in wild-type A549 cell lysates with no signal observed at this size in TGFBR2 knockout cell line. To generate this image, wild-type and TGFBR2 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 fluorescent western blot (TBS-based) blocking solution 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 40 µg

Lane 2:

TGFBR2 knockout A549 cell lysate at 40 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 65 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab283230, the same antibody clone in a different buffer formulation.

Western blot : Rabbit Monoclonal [EPR24349-124] to TGF beta Receptor II ab259360 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 70 kDa in Wild-type A549 cell lysates with no signal observed at this size in TGFBR2 knockout A549 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.

All lanes:

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human TGFBR2 (TGF beta Receptor II) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-tgfbr2-tgf-beta-receptor-ii-knockout-a549-cell-line-ab277895'>ab277895</a>) at 20 µg

Lane 2:

TGFBR2 knockout A549 at 20 µg

Secondary

All lanes:

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

Predicted band size: 65 kDa

Observed band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab259360, the same antibody clone in a different buffer formulation.

ab259360 was shown to react with TGFBR2 in wild-type A549 cells in Western blot with loss of signal observed in TGFBR2 knockout cell line ab261863. Wild-type A549 and TGFBR2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab259360 overnight at 4°C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 lysate at 30 µg

Lane 2:

TGFBR2 knock-out A549 lysate at 30 µg

Lane 2:

Western blot - Human TGFBR2 (TGF beta Receptor II) knockout A549 cell line (<a href='/en-us/products/cell-lines/human-tgfbr2-tgf-beta-receptor-ii-knockout-a549-cell-line-ab261863'>ab261863</a>)

Lane 2:

Western blot - Human TGFBR2 (TGF beta Receptor II) knockout A549 cell lysate (<a href='/en-us/products/cell-lysates/human-tgfbr2-tgf-beta-receptor-ii-knockout-a549-cell-lysate-ab261672'>ab261672</a>)

Observed band size: 66-95 kDa,48 kDa

false

• WB

Lab

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] - BSA and Azide free (AB283230)

This data was developed using ab259360, the same antibody clone in a different buffer formulation.

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

Negative control : Neuro-2a (PMID : 1310899).

Lysates used in this blot have experienced freeze-thaw cycles. Bands below 50 kDa are caused by degradation.

Exposure time : 81 seconds.

All lanes:

Western blot - Anti-TGF beta Receptor II antibody [EPR24349-124] (<a href='/en-us/products/primary-antibodies/tgf-beta-receptor-ii-antibody-epr24349-124-ab259360'>ab259360</a>) at 1/1000 dilution

Lane 1:

Mouse lung tissue lysate at 20 µg

Lane 2:

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

Lane 3:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) 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 1/100000 dilution

Predicted band size: 65 kDa

Observed band size: 70-80 kDa

false