Immunohistochemical analysis of paraffin-embedded human gastric cancer (B) and adjacent non-cancerous stomach tissue (A) labeling MRP1 with ab233383 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Membranous and cytoplasmic staining in human gastric cancer tissue (B) with weak staining in its adjacent non-cancerous tissue (A)
(PMID:23667609) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

All lanes : Anti-MRP1 antibody [EPR21062] (ab233383) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MRP1 knockout HeLa cell lysate
Lane 3 : Wild-type A549 cell lysate
Lane 4 : MRP1 knockout A549 cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 172 kDa

Lanes 1-4: Merged signal (red and green). Green - ab233383 observed at 250 kDa. Red - loading control ab7291 observed at 50 kDa.

 ab233383 Anti-MRP1 antibody [EPR21062] was shown to specifically react with MRP1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265256 (knockout cell lysate ab257242) was used. Wild-type and MRP1 knockout samples were subjected to SDS-PAGE. ab233383 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C 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.

 

 

ab233383 staining MRP1 in wild-type HeLa cells (top panel) and ABCC1 knockout HeLa cells (ab265256) (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab233383 at 1/100 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor^® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor^® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8).

All lanes : Anti-MRP1 antibody [EPR21062] (ab233383) at 1/1000 dilution

Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : A549 (human lung carcinoma cell line) whole cell lysate
Lane 3 : BxPC-3 (human pancreas adenocarcinoma cell line) whole cell lysate
Lane 4 : SK-OV-3 (human ovarian cancer cell line) whole cell lysate
Lane 5 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Developed using the ECL technique.

Predicted band size: 172 kDa
Observed band size: 250 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM/TBST.

The expression profile is consistent with what has been described in the literature (PMID: 22353810).

Immunohistochemical analysis of paraffin-embedded human lung cancer (B) and adjacent non-cancerous lung tissue (A) labeling MRP1 with ab233383 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Strong membranous and cytoplasmic staining in human lung cancer tissue (B) with weak staining in its adjacent noncancerous tissue (A) (PMID:23667609) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunohistochemical analysis of paraffin-embedded human esophagus cancer (B) and adjacent non-cancerous esophagus tissue (A) labeling MRP1 with ab233383 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Strong membranous and cytoplasmic staining in human esophageal cancer tissue (B) while staining is weak in its adjacent noncancerous tissue(A) (PMID: 26870278) is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

Immunofluorescent analysis of 100% methanol-fixed A549 (human lung carcinoma cell line) cells labeling MRP1 with ab233383 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in A549 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

 

Immunofluorescent analysis of 100% methanol-fixed BxPC-3 (human pancreas adenocarcinoma cell line) cells labeling MRP1 with ab233383 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in BxPC-3 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.