Solarbio乳酸去氫酶 (乳酸脫氫酶) 活性測定試劑盒

$60.00

運費美元 45 - 超過美元免費 300

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描述

Item Number: BC0685

規格: 100 Tests/48 Strips

介紹

  • The Lactate Dehydrogenase (乳酸脫氫酶) Activity Assay Kit quantifies the activity of LDH enzymes in biological samples.
  • LDH is crucial in cellular respiration, converting lactate to pyruvate.
  • Elevated LDH levels in tissues or body fluids may indicate cell damage or pathology.
  • The assay is utilized in research and clinical labs for diagnostic and experimental purposes.

內容

試劑名稱規格儲存條件
萃取液液體60 毫升× 1 瓶子
試劑一液體7 毫升× 1 瓶子
試劑二粉末1 Vial
試劑三液體7 毫升× 1 瓶子
試劑四液體25 毫升× 1 瓶子
Standard Solution液體1 毫升× 1 Vial
  • 試劑二: 使用前, 添加 1.3 mL of distilled water to fully dissolve. Once prepared, aliquot into small tubes and store at -20°C for up to 2 週. Avoid repeated freeze-thaw cycles.
  • Standard Solution: Sodium pyruvate solution with a concentration of 20 微摩爾/毫升.

Key Components of the LDH Activity Assay Kit:

  1. Substrate Solution: Contains the necessary components for the enzymatic reaction, typically including lactate and NAD^+ (nicotinamide adenine dinucleotide).
  2. Reaction Buffer: Provides optimal conditions for the enzymatic reaction.
  3. LDH Enzyme Standard: A reference standard with a known concentration of LDH activity for the generation of a standard curve.
  4. LDH Assay Reagent: A colorimetric or fluorometric reagent that detects the product of the LDH reaction, often leading to a color change or fluorescence.

How to Use LDH Activity Assay Kit:

The following is a general guide on how to use an LDH Activity Assay Kit. Specific protocols may vary between kits, and it’s crucial to follow the instructions provided by the kit manufacturer:

  1. 樣品製備: Prepare your biological samples (例如, cell lysates, tissue homogenates, or body fluids) and dilute them appropriately in a compatible buffer if needed.
  2. Standard Curve Preparation: Prepare a series of standards with known concentrations of LDH enzyme activity using the provided LDH enzyme standard. This is typically achieved by diluting the standard in a buffer or sample matrix.
  3. Assay Plate Setup: Add the diluted samples and standards to the wells of a microplate or cuvette.
  4. Addition of Substrate Solution: Add the substrate solution to each well or cuvette containing the samples and standards. The enzymatic reaction will convert lactate to pyruvate.
  5. 孵: Incubate the reaction mixture for a specified period, typically at a controlled temperature. The enzymatic reaction will occur, generating NADH (reduced form of NAD^+).
  6. 測量: Measure the absorbance or fluorescence of the reaction mixture at the appropriate wavelength using a microplate reader or fluorometer.
  7. 數據分析: Use the standard curve to convert absorbance or fluorescence values into LDH enzyme activity units.
  8. 品質管制: Check the precision and accuracy of your results by ensuring that the assay performance falls within the acceptable range.

實驗實例:

  1. Weighed 0.103g of Sedum leaves, added 1mL of extraction solution, homogenized in an ice bath, centrifuged at 8000g, 4°C 為 10 分分鐘, and took the supernatant for measurement on ice. 然後, following the assay steps, ΔA was calculated as A_test tubeA_control tube = 0.275 – 0.199 = 0.076. Using the standard curve equation y = 0.5218x + 0.0063 (R^2 = 0.9983), we obtained x = 0.134 微摩爾/毫升. Calculating lactate dehydrogenase (L-LDH) activity: L-LDH (U/克質量) = 66.67 × x ÷ W = 86.74 U/g
  2. Weighed 0.109g of rabbit liver, added 1mL of extraction solution, homogenized in an ice bath, centrifuged at 8000g, 4°C 為 10 分分鐘, and took the supernatant diluted 80 times with distilled water and placed it on ice for measurement. 然後, following the assay steps, ΔA was calculated as A_test tubeA_control tube = 0.795 – 0.132 = 0.663. Using the standard curve equation y = 0.5218x + 0.0063 (R^2 = 0.9983), we obtained x = 1.259 微摩爾/毫升. Calculating lactate dehydrogenase (L-LDH) activity: L-LDH (U/克質量) = 66.67 × x ÷ W × dilution factor = 61605.53 U/g
  3. Took 10μL of horse serum and followed the assay steps. Using the 96-well plate, ΔA was calculated as A_test tubeA_control tube = 0.415 – 0.161 = 0.254. Using the standard curve equation y = 0.5218x + 0.0063 (R^2 = 0.9983), we obtained x = 0.475 微摩爾/毫升. Calculating lactate dehydrogenase (L-LDH) activity: L-LDH (單位/毫升) = 66.67 × x = 31.67 U/m

附加資訊

重量0.65 公斤
尺寸

100T/48S

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