1,5-Anhydroglucitol Uptake Assay Kit (Cell-Based) (BN00908)

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BN00908
  • 1,5-Anhydroglucitol Uptake Assay Kit (Cell-Based) (BN00908)
  • 1,5-Anhydroglucitol Uptake Assay Kit (Cell-Based) (BN00908)
  • 1,5-Anhydroglucitol Uptake Assay Kit (Cell-Based) (BN00908)
$1,360.32

Description

ELISA Kit Technical ManualMSDS

1,5-Anhydroglucitol Uptake Assay Kit (Cell-Based)

1,5-Anhydroglucitol (1,5-AG) is the 1-deoxy form of D-glucose. Its level in blood plasma has been used in clinical research to investigate short-term glycemic control levels in diabetic patients. Cereals and animal source proteins constitute the main 1,5-AG source for humans. Most of 1,5-AG is absorbed by renal tubules, and its levels in human body are highly regulated and relatively consistent. Glucose can act as a competitive inhibitor of 1,5-AG in blood. Therefore, glucose levels are inversely proportional to 1,5-AG concentrations. The physiological function of 1,5-AG is not well understood. Recent studies showed 1,5-AG uptake could be associated with sodium-dependent glucose transporter. Assay Genie's 1,5-Anhydroglucitol Uptake Assay Kit utilizes a proprietary fluorescent 1,5-AG analog, which can be taken up by cells. However, this 1,5-AG analog cannot be fully utilized in glycolytic processes and thus accumulates inside the cells. Fluorescence generated by this analog is proportional to the cellular 1,5-AG uptake. The assay can be used to monitor 1,5-AG uptake by using fluorescent microscopy and flow cytometry. Phloretin, a compound that can inhibit glucose transporter and 1,5-AG cell uptake, is included in the kit as a control. This non-radioactive assay kit is easy-to-use and allows qualitative and quantitative measurements of 1,5-AG uptake in cultured cells

Figure: 1,5-AG uptake in Jurkat and HeLa cells. Jurkat cells (2.5x10 5 cells) were incubated in the presence or absence of Phloretin (4 µl, 100X concentration) for 45 min. After incubation time, cells were washed and incubated with Reagent, and the same concentration of Phloretin for another 30 min. according to kit’s protocol. (A) Flow Cytometry histograms comparing the inhibition of 1,5-AG uptake by Phloretin in Jurkat cells (Black: negative control cells; orange: Phloretin-treated cells; blue: untreated cells). (B) Fluorescence microscopy images of Jurkat cells. Top: Phloretin and 1,5-AG treated cells; Bottom: 1,5-AG treated cells. (C) 1,5-AG uptake in HeLa cells: HeLa cells showing the uptake of Reagent in the cytoplasm. Cells were stained with Reagent for 30 min. and fixed. Image was taken using a fluorescent microscope with a 60X oil objective lens.

Key Information Description

Product SKU

BN00908

Size

50 assays

Detection Method

FACS (488 nm excitation laser) and fluorescent microscope (excitation range 420 nm-495 nm)

Applications

  • Measurement of 1,5-Anhydroglucitol uptake in response to insulin, growth factors, cytokines, mitogens and nutrients, etc.
  • Dual-staining of 1,5-AG transporters and 1,5-AG uptake
  • Analysis of 1,5-AG metabolism and cell signaling in various cell typesScreening of anti-diabetic compounds

Features and Benefits

  • Easy-to-use
  • Non-radioactive
  • Image and accurately measure 1,5-Anhydroglucitol uptake in cultured cells in response to insulin, growth factors etc.

Kit Components

  • Analysis Buffer
  • AGTracker Reagent (100X)
  • Phloretin (100X) (in DMSO)
  • Phloretin (100X)

Storage Conditions

-20°C

Shipping Conditions

Gel Pack

Usage

For Research Use Only! Not For Use in Humans.

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