Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free (AB247826)

This data was developed using ab109301, the same antibody clone in a different buffer formulation.

ab109301 (purified) at 1/30 immunoprecipitating TR4 in 10 μg HEK293 cell lysate (Lanes 1 and 2, observed at 67 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration : 5% NFDM/TBST Dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-TR4 antibody [EPR1773(2)] (<a href='/en-us/products/primary-antibodies/tr4-antibody-epr17732-ab109301'>ab109301</a>)

Predicted band size: 65 kDa

false

• WB

Lab

Western blot - Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free (AB247826)

This data was developed using ab109301, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TR4 antibody [EPR1773(2)] (<a href='/en-us/products/primary-antibodies/tr4-antibody-epr17732-ab109301'>ab109301</a>) at 1/5000 dilution

All lanes:

HEK-293 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

Predicted band size: 65 kDa

Observed band size: 67 kDa

false

• WB

Lab

Western blot - Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free (AB247826)

This data was developed using ab109301, the same antibody clone in a different buffer formulation.

Lanes 1-3 : Merged signal (red and green). Green - ab109301 observed at 67 kDa. Red - loading control ab8245 observed at 36 kDa.

ab109301 Anti-TR4 antibody [EPR1773(2)] was shown to specifically react with TR4 in wild-type HEK293T cells. The band observed in CRISPR/Cas9 edited cell line ab266228 (CRISPR/Cas9 edited cell lysate ab257563) lane below 67 kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and TR4 CRISPR/Cas9 edited samples were subjected to SDS-PAGE. ab109301 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-TR4 antibody [EPR1773(2)] (<a href='/en-us/products/primary-antibodies/tr4-antibody-epr17732-ab109301'>ab109301</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

NR2C2 CRISPR/Cas9 edited HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human NR2C2 (TR4) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-nr2c2-tr4-knockout-hek-293t-cell-line-ab266228'>ab266228</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 65 kDa

Observed band size: 67 kDa

false

• WB

Unknown

Western blot - Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free (AB247826)

This data was developed using ab109301, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TR4 antibody [EPR1773(2)] (<a href='/en-us/products/primary-antibodies/tr4-antibody-epr17732-ab109301'>ab109301</a>) at 1/1000 dilution

Lane 1:

PC3 cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Predicted band size: 12 kDa,134 kDa,14 kDa,37 kDa,52 kDa,55 kDa,65 kDa,96 kDa

Observed band size: 14 kDa,67 kDa

false

• WB

Lab

Western blot - Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free (AB247826)

This data was developed using ab109301, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-TR4 antibody [EPR1773(2)] (<a href='/en-us/products/primary-antibodies/tr4-antibody-epr17732-ab109301'>ab109301</a>) at 1/5000 dilution

Lane 1:

PC-12 whole cell lysate at 20 µg

Lane 2:

3T3-L1 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

Predicted band size: 65 kDa

Observed band size: 67 kDa

false

• WB

Lab

Western blot - Anti-TR4 antibody [EPR1773(2)] - BSA and Azide free (AB247826)

This data was developed using ab109301, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TR4 antibody [EPR1773(2)] (<a href='/en-us/products/primary-antibodies/tr4-antibody-epr17732-ab109301'>ab109301</a>) at 1/2000 dilution

All lanes:

PC-3 whole cell lysate at 10 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

Predicted band size: 65 kDa

Observed band size: 67 kDa

false