{"product_id":"enzyfluo-ampk-phosphorylation-assay-kit-bht15600119","title":"EnzyFluo™ AMPK Phosphorylation Assay Kit","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eFor quantitative fluorescent immunoenzymatic assay of AMPK phosphorylation status in cultured cells. The assay uses Cell-Based ELISA (FL530\/585 nm, 360\/450 nm) for signal readout. Compatible sample input includes Cells. Typical stated assay timing is Assay takes 6.5 hrs, hands-on time 2.5 hrs.\u003c\/p\u003e\n\n\u003ch2\u003eKey elements and design rationale\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eReadout format:\u003c\/strong\u003e Cell-Based ELISA (FL530\/585 nm, 360\/450 nm) supports plate-based signal acquisition and consistent comparison across matched samples.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSample compatibility:\u003c\/strong\u003e The stated sample scope includes Cells, which is useful when aligning matrix type with calibration and control design.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eWorkflow timing:\u003c\/strong\u003e The listed assay time of Assay takes 6.5 hrs, hands-on time 2.5 hrs helps frame batch planning, replicate handling, and plate throughput.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eFeature emphasis:\u003c\/strong\u003e Sensitive. Can measure pAMPK modulation in as little as 500 cells\/well.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eAdditional feature notes highlight New and improved. Total assay time reduced from the standard 21 hours to 6.5 hours (hands-on time 2.5 hrs); Simple and convenient. Cells are directly cultured in 96-well plates. No cell lysis is necessary. Available format information for this listing includes 100 Tests.\u003c\/p\u003e\n\n\u003ch2\u003eBiological background\u003c\/h2\u003e\n\u003cp\u003eThis product is centered on measurement of enzyfluo ampk phosphorylation within the matrices described for the assay. In practice, datasets from this type of format are typically interpreted by comparing relative signal, activity, or abundance across matched control and experimental groups rather than relying on a single value in isolation. Careful alignment of sample matrix, incubation window, and calibration strategy is important when comparing results across plates, operators, or study days.\u003c\/p\u003e\n\n\u003ch2\u003eMore details\u003c\/h2\u003e\n\u003cp\u003eThe\u003ci\u003e 5-AMP-activated protein kinase \u003c\/i\u003e(AMPK) is a key sensor of intracellular energy balance. AMPK is activated in response to an increase in the AMP\/ATP ratio which can be caused by a number of factors such as muscle contraction, starvation, or hypoxia. AMPK is a heterotrimeric protein complex comprising α- (63 kDa), β- (38 kDa) and γ- (38 kDa) subunits. For each subunit, isoforms have been identified (α1, α2, β- 1, β- 2, γ1, γ2, γ3) which theoretically allow the formation of 12 different proteins. The α-subunit contains a serine\/threonine kinase domain and the regulatory subunits contain binding sites for AMP and ATP and for glycogen. AMPK is activated by phosphorylation on Thr-172 within the catalytic domain. AMP binding results in a 2 to 5-fold increase in AMPK activity compared to the basal level. The binding of AMP to the α-subunit causes allosteric activation of the kinase and induces a conformational change in the kinase domain that protects AMPK from dephosphorylation of Thr-172.BioAssay Systems cell-based ELISA measures phosphorylated AMPK in whole cells and normalizes the signal to the total protein content. The antibody recognizes both α-subunits and, thus, can be used for cells from all tissues (human, mouse, rat). This simple and efficient assay eliminates the need for cell lysate preparation and can be used to study AMPK regulation in short-term and long-term assays. In this assay, cells grown in 96-well plates are fixed and permeabilized in the wells. AMPK phosphorylation (pAMPK) is measured using a fluorescent ELISA followed by total protein measurement in each well.\u003cbr class=\"\"\u003e\u003cbr class=\"\"\u003e\u003c\/p\u003e\n\n\u003ch2\u003eDetection method\u003c\/h2\u003e\n\u003cp\u003eCell-Based ELISA (FL530\/585 nm, 360\/450 nm).\u003c\/p\u003e\n\n\u003ch2\u003eProcedures and timing\u003c\/h2\u003e\n\u003cp\u003eStated procedure or timing information: Assay takes 6.5 hrs, hands-on time 2.5 hrs.\u003c\/p\u003e\n\n\u003ch2\u003eResearch relevance and current trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003ePlate-based quantification and side-by-side group comparison remain central use cases for this assay format.\u003c\/li\u003e\n  \u003cli\u003eThe product notes emphasize multi-sample throughput, making it relevant for screening-oriented and larger batch comparison studies.\u003c\/li\u003e\n  \u003cli\u003eShort assay timing and plate compatibility support time-course or repeated-measure collection plans when handling is kept consistent.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eCommon research applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eQuantify enzyfluo ampk phosphorylation in cells by Cell-Based ELISA (FL530\/585 nm, 360\/450 nm) readout.\u003c\/li\u003e\n  \u003cli\u003eCompare treatment or phenotype groups using matched cells handling.\u003c\/li\u003e\n  \u003cli\u003eMonitor time-course or pre\/post changes in cells across study conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eInterpretation is usually strongest when signal changes are assessed alongside matrix-matched controls, replicate agreement, and the assay's stated analytical window.\u003c\/p\u003e\n\n\u003ch2\u003eNotes for experimental interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eMatrix composition, background signal, and sample handling can influence apparent response; compare like-with-like whenever possible.\u003c\/li\u003e\n  \u003cli\u003eUse appropriate blanks, controls, and replicate wells to distinguish biological differences from plate, reagent, or handling variability.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003c!-- Sources (internal):\n- Product Description column\n- Key Features column\n- More Details column\n- Method \/ Sample Type(s) \/ Assay Time \/ Detection Limit \/ Detection Method columns\n- Procedures column\n- Screening Services column\n--\u003e","brand":"BioAssay Systems","offers":[{"title":"100 Tests","offer_id":53238317121901,"sku":"EAMPK-100","price":589.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/EAMPKfig.jpg?v=1776668351","url":"https:\/\/www.ebiohippo.com\/products\/enzyfluo-ampk-phosphorylation-assay-kit-bht15600119","provider":"BioHippo","version":"1.0","type":"link"}