K607 100

Manufactured by Abcam

Sourced in United States

K607-100 is a product offered by Abcam. It is a lab equipment item, but a detailed description cannot be provided while maintaining an unbiased and factual approach without extrapolation or interpretation.

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18 protocols using k607 100

NADPH-Dependent LDHA and ATP Quantification

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Cells were subjected to EPB41L4A-AS1 overexpression and knockdown, cultured for 48 h, harvested, incubated in NADPH antibody for 1 h, washed with PBS, resuspended in growth medium, and immediately analyzed by flow cytometry (absorption spectrum [abs.] 488 nm/em. 562 nm). The total cellular LDHA in the medium supernatant (Biovision, K607-100) and ATP (Biovision, K354-100) concentration were determined by colorimetric assay.

Zhu Y., Liu Q., Liao M., Diao L., Wu T., Liao W., Wang Z., Li B., Zhang S., Wang S., Xie W., Jiang Y., Xu N., Zeng Y., Yang B.B, & Zhang Y. (2019). Overexpression of lncRNA EPB41L4A-AS1 Induces Metabolic Reprogramming in Trophoblast Cells and Placenta Tissue of Miscarriage. Molecular Therapy. Nucleic Acids, 18, 518-532.

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Measuring Cellular Pyruvate and Lactate

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Cellular contents of pyruvate were measured using kits purchased from BioVision (K609-100) following the manufacturers’ instructions. Cellular contents of lactate were measured using specific kits purchased from BioVision (K607-100), following the manufacturer's instructions.

Chow H.M., Cheng A., Song X., Swerdel M.R., Hart R.P, & Herrup K. (2019). ATM is activated by ATP depletion and modulates mitochondrial function through NRF1. The Journal of Cell Biology, 218(3), 909-928.

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Quantifying D- and L-Lactate in Staphylococcal Biofilms

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D- and L-lactate concentrations in supernatants from S. aureus WT and lactate mutant biofilm in vitro, mouse implant-associated tissue homogenates, and human synovial fluid were determined using D- and L-Lactate colorimetric assays according to the manufacturer’s instructions (Cat. #s K667–100 and K607–100, respectively; BioVision). Murine IL-10 levels in supernatants from MDSC- or macrophage-biofilm co-cultures and tissue homogenates were quantified using a Mouse Inflammation Cytometric Bead Array (CBA; Cat #552364, BD Biosciences) or sandwich ELISA (Mouse DuoSet; Cat. #DY417–05, R&D Systems). IL-10 in human synovial fluid samples and human monocyte-derived macrophages was measured by ELISA (Cat. #430604, BioLegend).

Heim C.E., Bosch M.E., Yamada K.J., Aldrich A.L., Chaudhari S.S., Klinkebiel D., Gries C.M., Alqarzaee A.A., Li Y., Thomas V.C., Seto E., Karpf A.R, & Kielian T. (2020). Lactate production by Staphylococcus aureus biofilm inhibits HDAC11 to reprogram the host immune response during persistent infection. Nature microbiology, 5(10), 1271-1284.

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Measuring Lactate Production in sEND.1 Cells

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sEND.1 cells were seeded in a 6 well plate. After overnight culture, cells were treated with or without IFNγ (100 ng/mL, 315-05, Peprotech) and AZD5363 (10 μM, HY-15431, MCE) for 12 h. The cells were then collected to detect lactate production using a lactate colorimetric/fluorometric assay kit (k607-100, Biovision, USA) while following the manufacturer's instructions.

Wang R., Ni C., Lou X., Zhang L., Wang L., Yao X., Duan X., Wan J., Li P, & Qin Z. (2023). IFNγ blockade in capillary leak site improves tumour chemotherapy by inhibiting lactate-induced endocytosis of vascular endothelial-cadherins. International Journal of Biological Sciences, 19(5), 1490-1508.

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Extracellular Lactate Quantification

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Extracellular lactate levels were measured using the Lactate Colorimetric/Fluorometric Assay kit according to the protocol suggested by the manufacturer (Biovision (Catalog # K607-100). Two microliters of culture medium were obtained from cells that were in culture for 24 hours. The absorbances (OD 570 nm) of the samples were measured using a BioTek Synergy 2 microplate reader.

Franklin D.A., He Y., Leslie P.L., Tikunov A.P., Fenger N., Macdonald J.M, & Zhang Y. (2016). p53 coordinates DNA repair with nucleotide synthesis by suppressing PFKFB3 expression and promoting the pentose phosphate pathway. Scientific Reports, 6, 38067.

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Comprehensive Serum Biochemistry Analysis

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To determine triglyceride, cholesterol, lactate and uric acid levels in blood, serum was collected from each mouse group. Total cholesterol and uric acid were analyzed using kits (TR13421 and TR24321) obtained from Thermo Fisher Scientific (Waltham, MA, USA). Triglycerides were measured with a Serum Triglyceride Determination Kit (TR0100, Sigma-Aldrich, St. Louis, MO, USA) and lactate measured with a colorimetric/fluorometric kit (K607–100, Biovision, Milpitas, CA, USA). Liver transaminase activity (AST/ALT) was analyzed using colorimetric kits (K753-100 and K752-100, Biovision). To determine serum biochemistry and liver transaminase levels, standard curves for each respective kit were prepared and samples were analyzed according to manufacturer’s instructions using the SpectraMax i3x (Molecular Devices, Sunnyvale, CA, USA).

Arends C.J., Wilson L.H., Estrella A., Kwon O.S., Weinstein D.A, & Lee Y.M. (2022). A Mouse Model of Glycogen Storage Disease Type IX-Beta: A Role for Phkb in Glycogenolysis. International Journal of Molecular Sciences, 23(17), 9944.

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Cytokine and Lactate Secretion Analysis

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HepG2 cells stably overexpressed control or serpinc1 were seeded in six-well plates (0.2×10⁶/well) for 24 h. Then, cells were washed and refreshed with 1 ml culture medium for 48 h. After centrifugation at 15,000 g for 10 min, the culture medium was collected and the ELISA kit was used to detect cytokines [including IL4 (Biolegend, 430301, San Diego, CA, USA), IL10 (Biolegend, 430601), and IL13 (Biolegend, 435207)] or lactate (BioVision, K607-100, Milpitas, CA, USA) according to the manufacturer’s protocols. The absorbance at 570 nm subtracted to 450 nm was estimated using a microplate reader (Multiskan GO, Thermo, Waltham, MA, USA).

Xu D., Wu J., Dong L., Luo W., Li L., Tang D, & Liu J. (2021). Serpinc1 Acts as a Tumor Suppressor in Hepatocellular Carcinoma Through Inducing Apoptosis and Blocking Macrophage Polarization in an Ubiquitin-Proteasome Manner. Frontiers in Oncology, 11, 738607.

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Metabolomic Analysis of MEFs

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For metabolomics analysis, MEFs were seeded on 100 mm plates, left to attach overnight and treated as indicated. Cell culture medium was removed and cells were rinsed with PBS and homogenized in acetonitrile, isopropanol and water (ratio of 3:3:2), and analyzed by automatic liner exchange/cold injection GC-TOF mass spectrometry at West Coast Metabolomics Centre (Davis, CA) for measurement of metabolites. For evaluation of cellular ammonia and lactate, MEFs were treated as indicated, conditioned media was collected and cells were rinsed once with PBS and snap frozen in liquid nitrogen. Extracellular and intracellular ammonia were quantified with an ammonia assay kit (Biovision #K370-100), and intracellular lactate was determined with a lactate assay kit (Biovision #K607-100) following the recommended manufacturer’s protocol.

Tan H.W., Sim A.Y, & Long Y.C. (2017). Glutamine metabolism regulates autophagy-dependent mTORC1 reactivation during amino acid starvation. Nature Communications, 8, 338.

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Lactate Colorimetric Assay in PDAC Cells

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Lactate production in PDAC cells was detected using the lactate colorimetric assay kit (K607-100, BioVision, Inc., Milpitas, CA, USA) in accordance with the manufacturer's instructions. Cells were seeded in a 96-well plate at a density of 1 × 10⁴ cells/well 1 day prior to the assay, which was performed in triplicate. In the assay, 5–20 μL of the sample was placed at a density of 50 μL/well on a 96-well plate containing the lactate assay buffer and incubated for 30 min at room temperature and away from light. Absorbance was measured using a microplate reader at 570 nm.

Park J.M., Peng J.M., Shen Y.S., Lin C.Y., Hsu T.W., Su Y.H., Chen H.A., Saengboonmee C., Chang J.S., Chiu C.F, & Shan Y.S. (2022). Phosphomimetic Dicer S1016E triggers a switch to glutamine metabolism in gemcitabine-resistant pancreatic cancer. Molecular Metabolism, 65, 101576.

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Osteoblast Differentiation and Lactate Measurement

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3T3-L1 and MC3T3-E1 cells were plated in 6-well plates at 2×10⁵ cells per well in 2 ml of the appropriate growth medium as above (section 2.1). After the cells reached confluence (about 1 d), we started osteoblast differentiation in half of the wells with the other half being the growth medium controls. The cells were differentiated for 7 (3T3-L1) or 14 days (MC3T3-E1) with complete medium changes every other day. On day 7 (3T3-L1) or day 14 (MC3T3-E1), cells were washed with 1X PBS then incubated in serum- and phenol red-free DMEM or α-MEM medium as appropriate, containing 25 mM glucose. Media were collected after 30 min incubation and lactate levels were measured using a colorimetric kit (Biovision, catalog # K607-100) following the manufacturer’s protocol. The values obtained were normalized to total protein content of the original wells.

Guntur A.R., Gerencser A.A., Le P.T., DeMambro V.E., Bornstein S.A., Mookerjee S.A., Maridas D.E., Clemmons D.E., Brand M.D, & Rosen C.J. (2018). Osteoblast like MC3T3-E1 cells prefer glycolysis for ATP production but adipocyte like 3T3-L1 cells prefer oxidative phosphorylation. Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, 33(6), 1052-1065.

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