Pkm2 d78a4

Manufactured by Cell Signaling Technology

Sourced in Germany

PKM2 (D78A4) is a recombinant monoclonal antibody that specifically recognizes pyruvate kinase M2 (PKM2). PKM2 is a key enzyme involved in glycolysis and energy metabolism in cells.

Automatically generated - may contain errors

Lab products found in correlation

Hspa8, by Novus Biologicals (1 mentions) Fitc conjugated secondary antibody, by BD (1 mentions) Anxa1, by R&D Systems (1 mentions) Aldehyde sulfate latex beads, by Thermo Fisher Scientific (1 mentions) Tsg101, by Abcam (1 mentions) Gallios flow cytometer, by Beckman Coulter (1 mentions) Bis tris gel, by Thermo Fisher Scientific (1 mentions) Iblot, by Thermo Fisher Scientific (1 mentions) Mitoprofile total oxphos human wb antibody cocktail, by Abcam (1 mentions) Odyssey blocking buffer, by LI COR (1 mentions) β tubulin, by Santa Cruz Biotechnology (1 mentions)

2 protocols using pkm2 d78a4

Exosome Flow Cytometry Analysis

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Exosome flow-cytometry analysis was performed as previously described¹⁵. Briefly, exosomes preparations (15 μg) were incubated with 10 μL of 4-μm-diameter aldehyde/sulfate latex beads (Life Technologies) for 15 min at RT, then resuspended in 1 mL PBS and incubated on a test tube rotator wheel overnight at 4 °C. Free binding sites on beads were saturated with 110 μl of 1 M glycine at RT for 30 min. Exosomes-coated beads were resuspended into 400 μL PBS containing 2% BSA and a 25 μL aliquot was incubated with the following antibodies: CD63 (sc-5275, Santa Cruz), CD9 (#655433, BD Bioscience), TSG101 (ab83, Abcam), ANXA1 (AF3770, R&D Biosystems), H2AFZ (sc-67218, Santa Cruz), HSPA8 (NB100-41377, Novus), PKM2 (D78A4, Cell Signaling), and anti-human IgG (#409309, BioLegend), or an isotype-matched negative control antibody for 30 min at 4 °C followed, when needed, by incubation with FITC-conjugated secondary antibody (BD Bioscience). One microgram of primary and secondary antibody was used for all stainings. Flow-cytometry data were acquired on a Gallios flow cytometer (Beckman Coulter) and analyzed using FlowJo.

Capello M., Vykoukal J.V., Katayama H., Bantis L.E., Wang H., Kundnani D.L., Aguilar-Bonavides C., Aguilar M., Tripathi S.C., Dhillon D.S., Momin A.A., Peters H., Katz M.H., Alvarez H., Bernard V., Ferri-Borgogno S., Brand R., Adler D.G., Firpo M.A., Mulvihill S.J., Molldrem J.J., Feng Z., Taguchi A., Maitra A, & Hanash S.M. (2019). Exosomes harbor B cell targets in pancreatic adenocarcinoma and exert decoy function against complement-mediated cytotoxicity. Nature Communications, 10, 254.

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Western Blot Analysis of Adipocyte Proteins

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Preadipocytes and adipocytes were lysed (30 min at 4 °C), cleared by centrifugation and protein concentrations were determined using BCA protein assay (Pierce). 15 or 30 μg protein lysate were separated on a 4–12% BisTris gel (Invitrogen) and blotted onto a Nitrocellulose Membrane using iBlot (Invitrogen). Membrane was blocked for 1 h in Odyssey Blocking Buffer (LiCor, Lincoln, NE USA) followed by incubation with primary antibodies. Subsequently, IRDye® or AlexaFluor® secondary antibodies (LiCor or Abcam, Cambridge, England) were used and signals were detected using the Odyssey Sa or classic (LiCor). Following antibodies were use: MitoProfile® Total OXPHOS Human WB Antibody Cocktail (#ab110411, abcam), PFKP (D4B2, Cell Signaling), PKM2 (D78A4, Cell Signaling) and β-tubulin (Santa Cruz, Heidelberg, Germany or Abcam).

Keuper M., Berti L., Raedle B., Sachs S., Böhm A., Fritsche L., Fritsche A., Häring H.U., Hrabě de Angelis M., Jastroch M., Hofmann S.M, & Staiger H. (2018). Preadipocytes of obese humans display gender-specific bioenergetic responses to glucose and insulin. Molecular Metabolism, 20, 28-37.

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