Anti-MRP1 antibody [EPR21062] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab233383).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab233383).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

This data was developed using the same antibody clone in a different buffer formulation (ab233383).

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).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab233383).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab233383).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab233383).

• WB

Lab

Western blot - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

This data was developed using ab234098, the same antibody clone in a different buffer formulation. Western blot : Rabbit Monoclonal [EPR21062] to MRP1 ab233383 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 172 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in abCC1 knockout MCF7 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-MRP1 antibody [EPR21062] (<a href='/en-us/products/primary-antibodies/mrp1-antibody-epr21062-ab233383'>ab233383</a>) at 1/1000 dilution

Lane 1:

Wild-type MCF7 at 20 µg

Lane 2:

ABCC1 knockout MCF7 at 20 µg

Lane 2:

Western blot - Human ABCC1 knockout MCF7 cell line (<a href='/en-us/products/cell-lines/human-abcc1-knockout-mcf7-cell-line-ab286378'>ab286378</a>) at 20 µg

Lane 3:

DU 145 at 20 µg

Lane 4:

Ramos at 20 µg

Secondary

All lanes:

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

Predicted band size: 172 kDa

Observed band size: 172 kDa

false

• WB

Lab

Western blot - Anti-MRP1 antibody [EPR21062] - BSA and Azide free (AB234098)

This data was developed using the same antibody clone in a different buffer formulation (ab233383).

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.

All lanes:

Western blot - Anti-MRP1 antibody [EPR21062] (<a href='/en-us/products/primary-antibodies/mrp1-antibody-epr21062-ab233383'>ab233383</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

MRP1 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

Western blot - Human ABCC1 (MRP1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-abcc1-mrp1-knockout-hela-cell-line-ab265256'>ab265256</a>)

Lane 3:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

MRP1 knockout A549 (Human lung carcinoma cell line) whole 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 1/20000 dilution

Predicted band size: 171 kDa

Observed band size: 250 kDa

false