Recombinant Anti-MRP1 antibody [EPR21061] (ab230948)

All lanes : Anti-MRP1 antibody [EPR21061] (ab230948) at 1/1000 dilution

Lane 1 : Wild-type A549 cell lysate
Lane 2 : ABCC1 knockout A549 cell lysate
Lane 3 : Wild-type HeLa cell lysate
Lane 4 : ABCC1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 171 kDa
Observed band size: 170 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab230948 observed at 170 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab230948 was shown to react with MRP1 in wild-type HeLa cells in Western blot with loss of signal observed in ABCC1 knockout cell line (ABCC1 knockout cell lysate). Wild-type HeLa and ABCC1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab230948 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 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.

Immunohistochemical analysis of paraffin-embedded human esophagus cancer (B) and its adjacent noncancerous tissue (A) labeling MRP1 with ab230948 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Strong membranous staining in human esophageal cancer tissue (B) with weak staining 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 a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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

All lanes : Anti-MRP1 antibody [EPR21061] (ab230948) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : A549 (human lung carcinoma epithelial cell) whole cell lysate
Lane 3 : BxPC-3 (human pancreas adenocarcinoma epithelial cell) whole cell lysate
Lane 4 : SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate
Lane 5 : HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 171 kDa


Exposure time: 3 minutes

Blocking/ Dilution buffer and concentration: 5% NFDM/TBST.

Immunohistochemical analysis of paraffin-embedded human lung cancer (B) and its adjacent noncancerous tissue (A) labeling MRP1 with ab230948 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). 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 a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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

Immunohistochemical analysis of paraffin-embedded human colon cancer (B) and its adjacent noncancerous tissue (A) labeling MRP1 with ab230948 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Strong membranous and cytoplasmic staining in human colon 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 a ready to use Goat Anti-Rabbit IgG H&L (HRP).

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