Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell pellet (B) MCT1 knockout HeLa cell pellet tissue labeling MCT1/Monocarboxylic acid transporter 1 with ab315382 at 1/10000 (0.049 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on (A) Wild-type HeLa (human cervix adenocarcinoma epithelial cell) cell pellet, negative staining on (B) MCT1 knockout HeLa cell pellet. The section was incubated with ab315382 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentThe knockout cell line was provided, with thanks, by RESOLUTE (re-solute.eu) Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling MCT1/Monocarboxylic acid transporter 1 with ab315382 at 1/10000 (0.049 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on human colon cancer (PMID : 27729975). The section was incubated with ab315382 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Immunohistochemical analysis of paraffin-embedded Human stomach cancer tissue labeling MCT1/Monocarboxylic acid transporter 1 with ab315382 at 1/10000 (0.049 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on human stomach cancer. The section was incubated with ab315382 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling MCT1/Monocarboxylic acid transporter 1 with ab315382 at 1/10000 (0.049 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Membranous staining on human stomach. The section was incubated with ab315382 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MCT1 knockout HeLa cells (MCT1 knockout human cervical adenocarcinoma epithelial cell) cells labelling MCT1/Monocarboxylic acid transporter 1 with ab315382 at 1/1000 (0.485 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in parental HeLa cells, and no staining in MCT1 knockout HeLa cells. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). The knockout cell line was provided, with thanks, by RESOLUTE (re-solute.eu)

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Flow cytometry overlay histogram showing wild-type A-549 (green line) and SLC16A1 knockout A-549 stained with ab315382 (magenta line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab315382) (1x 10⁶ in 100μl at 0.008 μg/ml (1/60625)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C.

Isotype control antibody was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control in A-549 WT cells (black line) and A-549-SLC16A1 KO cells (grey line), at the same conditions as the primary antibody.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

This antibody gave a positive signal in wild-type A-549 fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

• IP

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Immunoprecipitation - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

MCT1/Monocarboxylic acid transporter 1 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab315382 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315382 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

All lanes:

Immunoprecipitation - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (ab315382) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 2:

ab315382 at 1/30 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315382 in HeLa whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 6s

• WB

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Western blot - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 15505343).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, ab315382 was shown to bind specifically to MCT1/Monocarboxylic acid transporter 1. A band was observed at 43 kDa in wild-type HeLa cell lysates whereas no signal was observed at this size in the MCT1 knockout cell line.

The knockout cell line was provided, with thanks, by RESOLUTE (re-solute.eu).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane1-5 : 15 seconds; Lane 6 : 158 seconds

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

All lanes:

Western blot - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (ab315382) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

MCT1/Monocarboxylic acid transporter 1 knockout HeLa whole cell lysate at 20 µg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 6:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 43 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Western blot : Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] ab315382 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 43 kDa in Wild-type A549 cell lysates with no signal observed at this size in SLC16A1 knockout A549 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-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (ab315382) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

Western blot - Human SLC16A1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-slc16a1-knockout-a549-cell-line-ab301232'>ab301232</a>) at 20 µg

Secondary

All lanes:

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

Predicted band size: 54 kDa

Observed band size: 43 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] (AB315382)

Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling MCT1/Monocarboxylic acid transporter 1 with ab315382 at 1/10000 (0.049 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : No staining on human pancreas (PMID : 32355273). The section was incubated with ab315382 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.