Anti-TMUB1 antibody [EPR14066] - BSA and Azide free

• WB

Lab

Western blot - Anti-TMUB1 antibody [EPR14066] - BSA and Azide free (AB250234)

This data was developed using the same antibody clone in a different buffer formulation (ab180586).

Lanes 1- 3 : Merged signal (red and green). Green - ab180586 observed at 27 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab180586 was shown to react with TMUB1 in wild-type HeLa (Human epithelial line from cervix adenocarcinoma) cells in western blot. Loss of signal was observed when knockout cell line ab265852 (knockout cell lysate ab258237) was used. Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) and TMUB1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab180586 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 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-TMUB1 antibody [EPR14066] (<a href='/en-us/products/primary-antibodies/tmub1-antibody-epr14066-ab180586'>ab180586</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 2:

TMUB1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg

Lane 2:

Western blot - Human TMUB1 partial knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-tmub1-partial-knockout-hela-cell-line-ab265852'>ab265852</a>)

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate cell lysate at 20 µg

Predicted band size: 26 kDa

Observed band size: 27 kDa

false

• WB

Supplier Data

Western blot - Anti-TMUB1 antibody [EPR14066] - BSA and Azide free (AB250234)

This data was developed using ab180586, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-TMUB1 antibody [EPR14066] (<a href='/en-us/products/primary-antibodies/tmub1-antibody-epr14066-ab180586'>ab180586</a>) at 1/50000 dilution

Lane 1:

Human fetal liver tissue lysate at 20 µg

Lane 2:

Human cerebellum tissue lysate at 20 µg

Lane 3:

HepG2 cell line lysate lysate at 20 µg

Lane 4:

HeLa cell line lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 26 kDa

Observed band size: 24 kDa,27 kDa

false

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-TMUB1 antibody [EPR14066] - BSA and Azide free (AB250234)

This data was developed using ab180586, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde fixed HepG2 cells labeling TMUB1 with ab180586 at 1/500 (green). Dapi staining shown in blue.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-TMUB1 antibody [EPR14066] - BSA and Azide free (AB250234)

This data was developed using ab180586, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 2% paraformaldehyde fixed HeLa cells labeling TMUB1 with ab180586 at 1/70 (pink). Control shown in green.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TMUB1 antibody [EPR14066] - BSA and Azide free (AB250234)

This data was developed using ab180586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin embedded Human liver tissue labeling TMUB1 with ab180586 at 1/100.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-TMUB1 antibody [EPR14066] - BSA and Azide free (AB250234)

This data was developed using the same antibody clone in a different buffer formulation (ab180586).

Lanes 1 - 4 : Merged signal (red and green). Green - ab180586 observed at 27 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab180586 was shown to react with TMUB1 in wild-type HeLa cells in Western blot with loss of signal observed in TMUB1 knockout cell line ab278130 (knockout cell lysate ab278185). Wild-type HeLa and TMUB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab180586 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-TMUB1 antibody [EPR14066] (<a href='/en-us/products/primary-antibodies/tmub1-antibody-epr14066-ab180586'>ab180586</a>) at 1/10000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

TMUB1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human TMUB1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-tmub1-knockout-hela-cell-line-ab278130'>ab278130</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Human Cerebellum tissue lysate at 20 µg

Predicted band size: 26 kDa

Observed band size: 27 kDa

false