SLC16A1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
FLJ36745, HHF7, MCT, MCT 1, MGC44475, MOT1_HUMAN, Monocarboxylate transporter, Monocarboxylate transporter 1, Monocarboxylate transporter isoform 1, Monocarboxylic acid transporter 1, SLC16A1 protein, Slc16a1, Solute carrier family 16 (monocarboxylic acid transporters) member 1, Solute carrier family 16 member 1, Solute carrier family 16 member 1 (monocarboxylic acid transporter 1)
SLC16A1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (ab288558). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
MCT1 also known as Monocarboxylate Transporter 1 or SLC16A1 is a protein that facilitates the transport of monocarboxylates such as lactate and pyruvate across cell membranes. The approximate molecular weight of MCT1 is 51 kDa. This transporter expresses in many tissues including muscle brain liver and kidney. Its widespread presence indicates a significant role in cellular metabolism and energy balance across different biological systems.
MCT1 acts as an important transport mechanism for lactate and pyruvate driving the cellular uptake and release processes of these molecules. It partners with CD147 also known as Basigin to form a functional heteromeric complex. This binding allows the correct localization and stabilization of MCT1 in the plasma membrane ensuring effective transport activity. The presence of MCT1 in metabolically active tissues highlights its role in maintaining homeostasis in energy-rich environments.
MCT1 plays a significant role in the glycolysis and gluconeogenesis pathways by mediating lactate transport which connects anaerobic and aerobic metabolic processes. Through these pathways MCT1 associates with key metabolic enzymes like lactate dehydrogenase (LDH) that regulate the conversion between lactate and pyruvate. This association provides a link between lactate production and consumption thereby controlling pH levels and preventing the buildup of metabolic intermediates that can disrupt cellular functions.
MCT1 is implicated in cancer and metabolic acidosis. In cancer elevated expression of MCT1 supports the Warburg effect a phenomenon where cancer cells rely on aerobic glycolysis leading to increased lactate production. MCT1 works alongside proteins such as CD147 influencing tumor growth and metastasis by promoting lactate export. In metabolic acidosis impaired MCT1 function can disrupt lactate clearance affecting tissue pH balance and contributes to conditions like lactic acidosis. Understanding MCT1's role in these diseases highlights the importance of targeting its function for therapeutic purposes.
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Western blot: Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] Anti-MCT1/Monocarboxylic acid transporter 1 antibody [EPR26702-83] ab315382 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (Anti-Calnexin antibody [CANX/1543] 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] (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 (ab301232) at 20 µg
All lanes: Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 43 kDa
34 bp deletion after Leu 160 (allele 1); 15 bp deletion after Pro 162 (allele 2)
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