Anti-BOP1 antibody [6H11] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BOP1 antibody [6H11] - BSA and Azide free (AB255775)

This data was produced using the same clone but in a different formulation, ab252819.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (human chronic myelogenous leukemia cell line from bone marrow) cells labelling BOP1 with ab252819 at 1/50 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in K562 cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab252819 at a 1/50 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2 : ab179513 at a 1/200 dilution followed by ab150157 at a 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-BOP1 antibody [6H11] - BSA and Azide free (AB255775)

This data was produced using the same clone but in a different formulation, ab252819.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labelling BOP1 with ab252819 at 1/50 dilution, followed by ab150157 Goat Anti-Rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HeLa cells. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Negative control 1 : ab252819 at a 1/50 dilution followed by ab150080 at a 1/1000 dilution.

Negative control 2 : ab179513 at a 1/200 dilution followed by ab150157 at a 1/1000 dilution.

• IP

Lab

Immunoprecipitation - Anti-BOP1 antibody [6H11] - BSA and Azide free (AB255775)

This data was produced using the same clone but in a different formulation, ab252819.

BOP1 was immunoprecipitated from 0.35 mg K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab252819 at 1/30 dilution (2 μg in 0.35 mg lysates). Western blot was performed on the immunoprecipitate using ab252819 at 1/1,000 dilution. Goat Anti-Rat IgG (H+L), HRP (ab205720) was used at 1/10,000 dilution.

Lane 1 : K562 whole cell lysate 10 μg.

Lane 2 : ab252819 IP in K562 whole cell lysate.

Lane 3 : Rat IgG1 (ab18407) instead of ab252819 in K562 whole cell lysate.

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

Exposure time : 3 seconds.

All lanes:

Immunoprecipitation - Anti-BOP1 antibody [6H11] (<a href='/en-us/products/primary-antibodies/bop1-antibody-6h11-ab252819'>ab252819</a>)

Predicted band size: 84 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-BOP1 antibody [6H11] - BSA and Azide free (AB255775)

This data was produced using the same clone but in a different formulation, ab252819.

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 10 seconds; Lane 2 : 59 seconds; Lane 3 : 6 seconds.

All lanes:

Western blot - Anti-BOP1 antibody [6H11] (<a href='/en-us/products/primary-antibodies/bop1-antibody-6h11-ab252819'>ab252819</a>) at 1/5000 dilution

Lane 1:

Caco-2 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 3:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Predicted band size: 84 kDa

Observed band size: 110 kDa

false