糖化血红蛋白 (glycated hemoglobin,GHb) 是红细胞中的血红蛋白与血中糖类物质 (主要是葡萄糖) 缓慢、连续的非酶促反应的产物，一旦形成稳定的 GHb 则不再解离，在红细胞生存的 120 天内，其合成速率与红细胞所处环境中糖的浓度成正比，因此 GHb 可反映测定前较长时间内 (约 6~8 周) 血糖的平均水平，不受血糖暂时波动的影响。GHb 的检测方法较多，主要有离子交换层析法、亲和层析法、高效液相色谱法、免疫学方法、电泳法、比色法和离子捕获法等。糖化血红蛋白溶血剂由磷酸盐、表面活性剂等组成，主要用于进行糖化血红蛋白检测时的血液样本前处理，采用离子交换层析法测定 GHb 占总 Hb 的百分率。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
自备材料：
1、生理盐水
2、水浴锅、离心机、离心管
操作步骤 (仅供参考)：
1、溶血液的制备：将 EDTA 抗凝血或毛细管血 20 μL，加于 2 mL 生理盐水中，摇匀，离心，吸除上清，留红细胞沉淀，加入溶血剂 0.3 mL，摇匀，37℃ 水浴 15 min，除去不稳定的糖化血红蛋白 (不稳定 HbA1c)。
2、用离子交换层析法完成 GHb 百分率的测定。
注意事项：
1、溶血剂恢复至室温后使用，如有必要可分装使用。
2、为了您的安全和健康，请穿实验服并戴一次性手套操作。
3、试剂开封后请尽快使用，以防影响后续实验效果。
Glycated hemoglobin (GHb) is the product of a slow, continuous non-enzymatic reaction between hemoglobin in red blood cells and carbohydrates in the blood (primarily glucose). Once stable GHb is formed, it will not dissociate. Within the 120-day lifespan of red blood cells, its synthesis rate is proportional to the glucose concentration of the surrounding environment. Therefore, GHb can reflect the average blood glucose level over a relatively long period (approximately 6–8 weeks) prior to measurement and is not affected by temporary fluctuations in blood glucose. There are multiple detection methods for GHb, including ion-exchange chromatography, affinity chromatography, high-performance liquid chromatography, immunoassays, electrophoresis, colorimetry and ion capture assay. Glycated Hemoglobin Lysing Reagent consists of phosphates, surfactants and other components. It is mainly used for blood sample pretreatment in glycated hemoglobin detection, and ion-exchange chromatography is adopted to determine the percentage of GHb in total Hb. This reagent is for research use only and is not intended for clinical diagnosis or other applications.
Materials to be prepared by user:
1. Normal saline
2. Water bath, centrifuge, centrifuge tubes
Operating Procedure (For Reference Only):
1. Preparation of hemolysate: Add 20 μL of EDTA-anticoagulated blood or capillary blood into 2 mL of normal saline, mix well and centrifuge. Discard the supernatant and retain the red blood cell pellet. Add 0.3 mL hemolyzing reagent, mix thoroughly and incubate in a 37 ℃ water bath for 15 min to remove labile glycated hemoglobin (labile HbA1c).
2. Complete the determination of GHb percentage via ion-exchange chromatography.
Precautions:
1. Allow the hemolyzing reagent to warm to room temperature before use; aliquot if necessary.
2. For your safety and health, wear a lab coat and disposable gloves during operation.
3. Use the reagent promptly after opening to avoid adverse impacts on subsequent experimental results.
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