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SLC16A3 / MCT 4 Recombinant Rabbit Monoclonal Antibody [PSH06-91]

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Specification

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Catalog# HA722766

SLC16A3 / MCT 4 Recombinant Rabbit Monoclonal Antibody [PSH06-91]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Green monkey

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

SLC16A3 / MCT 4 Recombinant Rabbit Monoclonal Antibody [PSH06-91]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human SLC16A3 aa 385-465.

Species Reactivity

Human, Green monkey

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 50 kDa

Positive Control

HeLa cell lysate, HepG2 cell lysate, HCT 116 cell lysate, NCI-H441 cell lysate, COS-1 cell lysate, HeLa, human placenta tissue, human rectum tissue, human skeletal muscle tissue.

Conjugation

unconjugated

Clone Number

PSH06-91

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:20,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:1,000

  • FC

  • 1:1,000

Target

Function

Monocarboxylate transporter 4 (MCT4) also known as solute carrier family 16 member 3 is a protein that in humans is encoded by the SLC16A3 gene. Northern and western blotting and EST database analyses showed MCT4 to be widely expressed and especially so in glycolytic tissues such as white skeletal muscle fibers, astrocytes, white blood cells, chondrocytes, and some mammalian cell lines. Because of this, it has been proposed that the properties of MCT4 might be especially appropriate for export of lactate derived from glycolysis. MCT4 exhibits a lower affinity for most substrates and inhibitors than MCT1, with Km and Ki values some 5–10-fold higher. The high Km for pyruvate may be especially significant as this avoids loss of pyruvate from the cell which, were it to occur, would prevent removal of the reduced form of nicotinamide adenine dinucleotide (NADH) produced in glycolysis by reduction of pyruvate to lactate. MCT4 can be upregulated by HIF-1α and AMPK.

Background References

1. Monsorno K et al. Loss of microglial MCT4 leads to defective synaptic pruning and anxiety-like behavior in mice. Nat Commun. 2023 Sep

2. Babl N et al. MCT4 blockade increases the efficacy of immune checkpoint blockade. J Immunother Cancer. 2023 Oct

Subcellular Location

Cell membrane, Basolateral cell membrane.

UNIPROT

SwissProt: O15427 Human

Synonyms

MCT 4 antibody

MCT3 antibody

MCT4 antibody

Monocarboxylate transporter 3 antibody

Monocarboxylate transporter 4 antibody

MOT4_HUMAN antibody

SLC16A3 antibody

Solute carrier family 16 member 3 antibody

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of SLC16A3 / MCT 4 on different lysates with Rabbit anti-SLC16A3 / MCT 4 antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/20,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: K-562 cell lysate (negative)<br />Lane 4: HCT 116 cell lysate<br />Lane 5: NCI-H441 cell lysate<br />Lane 6: COS-1 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 50 kDa<br />Observed band size: 40 kDa<br /><br />Exposure time: 6 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of SLC16A3 / MCT 4 on different lysates with Rabbit anti-SLC16A3 / MCT 4 antibody (HA722766) at 1/20,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: K-562 cell lysate (negative)
    Lane 4: HCT 116 cell lysate
    Lane 5: NCI-H441 cell lysate
    Lane 6: COS-1 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 50 kDa
    Observed band size: 40 kDa

    Exposure time: 6 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722766) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling SLC16A3 / MCT 4 with Rabbit anti-SLC16A3 / MCT 4 antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SLC16A3 / MCT 4 antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling SLC16A3 / MCT 4 with Rabbit anti-SLC16A3 / MCT 4 antibody (HA722766) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-SLC16A3 / MCT 4 antibody (HA722766) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-SLC16A3 / MCT 4 antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-SLC16A3 / MCT 4 antibody (HA722766) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722766) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human rectum tissue with Rabbit anti-SLC16A3 / MCT 4 antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human rectum tissue with Rabbit anti-SLC16A3 / MCT 4 antibody (HA722766) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722766) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-SLC16A3 / MCT 4 antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue with Rabbit anti-SLC16A3 / MCT 4 antibody (HA722766) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA722766) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of HeLa cells labeling SLC16A3 / MCT 4.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722766" style="font-weight: bold;text-decoration: underline;">HA722766</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of HeLa cells labeling SLC16A3 / MCT 4.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722766, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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