本试剂盒采用双抗体夹心ELISA法检测样品中人CRP的浓度。人CRP捕获抗体已经预包被于酶标板上，当加入样品或标准品时，其中的人CRP会与捕获抗体结合，而其它游离成分则会通过洗涤被除去。接着，再加入生物素标记的人CRP抗体后，抗人CRP抗体与人CRP接合，形成夹心的免疫复合物，其它游离成分则通过洗涤被除去。随后加入酶复合物，生物素与酶复合物特异性结合，这样酶复合物上的HRP就与夹心的免疫复合物连接起来，而其它游离成分则通过洗涤被除去。最后加入显色剂，若样品中存在人CRP，则会形成免疫复合物，其上连接的HRP会催化无色的显色剂氧化生成蓝色物质，而后加入终止液，最终产物呈黄色。通过酶标仪检测，读取450 nm处的OD值，人CRP浓度与OD₄₅₀值之间呈正比，通过检测标准品绘制标准曲线，对照未知样品中OD值，即可计算出样品中人CRP的浓度。
背景简介
C-反应蛋白 (C-reactive protein, 简称CRP) 是一种在机体受到感染或组织损伤时血浆中急剧上升的蛋白质，也称为急性蛋白。CRP可以激活补体和加强吞噬细胞的吞噬作用，清除入侵机体的病原微生物和损伤、坏死、凋亡的组织细胞，在机体的天然免疫过程中发挥重要的保护作用。
CRP与磷脂胆碱残基具有高度亲和力，并且可以和多种自身配体或外来配体相结合，激活补体活化的经典途径，但限制补体激活晚期炎症反应的发展及强度。CRP还具有和IgG及补体相似的调理和凝集作用，增强巨噬细胞对各种细菌和异物的吞噬功能，从而减少由于外来抗原暴露所带来的异常免疫反应。
试剂盒组成
| 项目号 | 物理外观 | 组件 | 96T | 储存 | | H1509775A | — | 预包被酶标板 | 96孔 | 2-8℃. | | H1509775B | 液体 | 样品稀释液 | 30 mL | 2-8℃. | | H1509775C | 固体 | 重组人CRP标准品 (冻干) | 2 支 (50 ng/支) | 2-8℃. | | H1509775D | 液体 | 生物素标记人CRP抗体 | 130 μL (效价1:100) | 2-8℃. | | H1509775E | 液体 | 抗体稀释液 | 12 mL | 2-8℃. | | H1509775F | 液体 | 酶复合物 (HRP标记的链霉亲和素) | 130 μL (效价1:100) | 2-8℃. | | H1509775G | 液体 | 酶复合物稀释液 | 12 mL | 2-8℃. | | H1509775H | 液体 | 浓缩洗涤液 (25×) | 30 mL | 2-8℃. | | H1509775I | 液体 | 底物 (TMB) | 10 mL | 2-8℃. | | H1509775J | 液体 | 终止液 | 10 mL | 2-8℃. | | H1509775K | — | 封板膜 | 4张 | 2-8℃. |
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操作步骤
1. 样品制备
(1) 根据样品种类选择相应的处理方法：
A. 细胞上清：将细胞培养上清液100~500×g离心5 min，去除悬浮物后即可；
B. 血清样品：将全血在室温下静置0.5~2 h，待其自然凝固并析出血清后，离心取黄色上清即可 (4℃, 1000~2000×g, 10 min)，注意请勿吸取沉淀，制备好的血清需置于冰上待用，请勿在其中添加任何防腐剂或抗凝剂；
C. 血浆样品：使用EDTA对全血进行抗凝处理后，混合均匀置于冰上，离心取黄色上清即可 (4℃, 1000~2000×g, 10 min)，注意请勿吸取沉淀，制备好的血浆需置于冰上待用；
D. 组织匀浆/体液：离心去除沉淀即可。
注意：① 若待测样品无法及时检测，样品制备完成后，请分装冻存于-20℃，避免反复冻融；
② 请保证待测样品清澈透明，检测前如发现样品中有悬浮物，需通过离心去除；
③ 为了保证检测结果准确，请勿使用溶血、黄疸、高血脂或污染的样品。
(2) 稀释样品
查阅相关文献，预估样品中待测因子的含量，从而确定适当的稀释倍数，使稀释后样品中待测因子的浓度处于 ELISA 试剂盒的最佳检测范围。根据待测因子含量的不同，分别采取不同的稀释方案：
① 待测因子含量在500~5000 ng/mL 范围内，一般按1:100稀释，即向297 μL样品稀释液中加入3 μL样品；
② 待测因子含量在50~500 ng/mL范围内，一般按1: 10稀释，即向225 μL样品稀释液中加入25 μL样品；
③ 待测因子含量在0.78~50 ng/mL范围内，一般按1: 2稀释，即向100 μL样品稀释液中加入100 μL样品；
④ 待测因子含量≤0.78 ng/mL，样品一般无需稀释。
以上方案仅供参考，实验中请详细记录样品的稀释方法。
2. 检测准备工作
(1) 试剂盒自 4℃冰箱取出后，请置于室温平衡20 min；如从-20℃取出，各组分需彻底融化后再平衡20 min；检测完成后，剩余试剂请及时置于4℃或-20℃保存；
(2) 将浓缩洗涤液 (25×)用双蒸水或去离子水稀释成1×洗涤液；
(3) 重组人CRP标准品的稀释和使用 (在使用前2 h内准备，室温操作，请严格控制在25~28℃)
① 配制50 ng/mL标准品：取1 mL样品稀释液加入标准品管内，盖好后静置15 min以上，然后反复颠倒/搓动以助溶解；
② 按下表将50 ng/mL标准品用样品稀释液进行倍比梯度稀释。(最高浓度为50 ng/mL，将标准品稀释液作为浓度0 ng/mL。)
| 管号 | 稀释液用量 (μL) | 复溶后标准品用量 (μL) | 标准品的最终浓度 (ng/mL) | | A | 0 | 1,000 | 50
| | B | 300 | 300 (从A管中取) | 25
| | C | 300 | 300 (从B管中取) | 12.5
| | D | 300 | 300 (从C管中取) | 6.25
| | E | 300 | 300 (从D管中取) | 3.12
| | F | 300 | 300 (从E管中取) | 1.56
| | G | 300 | 300 (从F管中取) | 0.78
| | H | 300 | 0 | 0 |
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注意：标准品复溶加样后，剩余部分请丢弃。
(4) 准备生物素标记人CRP抗体工作液
① 按每孔需添加100 μL抗体工作液，计算其总用量 (为弥补操作中的损耗，需多配制100~200 μL)；
② 按1 μL生物素标记人CRP抗体添加99 μL抗体稀释液的比例配制工作液，轻轻混匀。
(5) 准备酶复合物工作液 (需在使用前1 h内准备)
① 按每孔需添加100 μL酶复合物工作液，计算其总用量 (为弥补操作中的损耗，需多配制100~200 μL）；
② 按1 μL酶复合物添加99 μL酶复合物稀释液的比例配制工作液，轻轻混匀。
3. 检测流程
(1) 通过计算确定一次实验所需的板条数，取出所需板条放置于框架内，多余的板条请放回铝箔袋密封，保存于4℃或-20℃;
注意：① 标准品和样品建议做双复孔检测；
② 每次实验均需绘制标准曲线。
(2) 将用样品稀释液稀释后的样品和不同浓度标准品 (100 μL/孔)分别加入相应孔中，用封板胶纸封住反应孔，37℃孵育90 min；
注意：① 请查阅相关文献确定样品中待检测蛋白的大致浓度，若其大于本试剂盒标准曲线的最大标准品浓度，请将样品适当稀释后再进行检测；
② 整个加样过程不宜超过10 min，否则可能会影响检测结果。
(3) 甩去酶标板内液体，无需洗板，将板倒扣在吸水纸上拍干；
(4) 加入稀释后的生物素标记人CRP抗体工作液 (100 μL/孔)，用封板胶纸封住反应孔，37℃孵育60 min；
(5) 洗板5次，每孔1×洗涤液用量为300 μL，注入与吸出间隔15~30 s，洗完后将板倒扣在吸水纸上拍干；
注意：洗涤过程至关重要，洗涤不充分会导致结果产生较大误差。
(6) 加入稀释后的酶复合物 (100 μL/孔)，用封板胶纸封住反应孔，37℃避光孵育30 min；
(7) 洗板5次，方法同步骤12；
(8) 加入显色剂TMB (100 μL/孔 )，用封板胶纸封住反应孔，避光37℃反应10~25 min；
注意：① 在保存和使用时，请勿将TMB接触氧化剂和金属；
② 因实验室条件差异，最佳显色时间会有所不同，反应充分时肉眼可见标准品的前3~4孔有明显的梯度蓝色。
(9) 加入终止液 (100 μL/孔 )，混匀后即刻使用酶标仪测量OD₄₅₀，同时设定540 nm或570 nm作为校正波长，即可计算得到校正吸光度值 (OD₄₅₀-OD₅₄₀或OD₄₅₀-OD₅₇₀)；
注意：读取OD值建议在10 min内完成。
4. 数据分析
(1) 绘制标准曲线。以标准品浓度作横坐标，OD值作纵坐标，利用计算机软件作四参数逻辑 (4-PL) 曲线拟合创建标准曲线，通过样品的OD值即可在标准曲线上计算出其相应浓度。
注意：① 复孔OD值在20%的差异范围内结果才有效，复孔OD值取平均后可作为测量值；
② 若样品OD值高于标准曲线上限，应适当稀释后重测，计算浓度时应乘以稀释倍数。
(2) 典型数据
该标准曲线仅供演示使用。每批检测样本均需单独绘制标准曲线。
| 标准品浓度 (ng/ml) | O.D. | | 0 | 0.087 | | 0.78 | 0.181 | | 1.56 | 0.253 | | 3.12 | 0.457 | | 6.25 | 0.826 | | 12.5 | 1.283 | | 25 | 1.857 | | 50 | 2.316 |
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(3) 精密度
① 板内精密度 (同批次检测精密度)：取3份已知浓度样本，在同一块酶标板上重复检测20次，用以评估板内精密度。
② 板间精密度 (批次间检测精密度)：取3份已知浓度样本，进行独立批次检测，用以评估板间精密度。实验由至少三名实验人员、使用两个不同批号的试剂盒组分完成。
| 板内精密度 | 板内精密度 | 板内精密度 | 板间精密度 | 板间精密度 | 板间精密度 | | 样本编号 | 1 | 2 | 3 | 1 | 2 | 3 | | 检测数量 (n) | 20 | 20 | 20 | 20 | 20 | 20 | | 平均值 (ng/ml) | 64
| 73
| 105
| 84
| 49
| 193
| | 标准差 | 5.1
| 2.5
| 5.4
| 2.6
| 1.8
| 8.9
| | 变异系数 (%) | 7.9
| 3.4
| 5.1
| 3.1
| 3.7
| 4.6
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(4) 回收率
在不同基质样本中，于检测量程范围内加入高、中、低三个浓度梯度的人C反应蛋白/CRP，对样本加标回收率进行评估。
| 样本类型 | 平均回收率 (%) | 回收率范围 | | 细胞培养上清液 (n=10) | 113
| 90%-119%
| | 血清 (n=10) | 102
| 88%-110%
| | 肝素血浆 (n=10) | 97
| 90%-118%
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(5) 线性关系
为了评估该检测方法的线性，将含有和/或添加了高浓度人C反应蛋白/CRP的样本，使用校准品稀释液进行系列稀释，从而制备出数值落在该检测方法动态范围内的样本。
| 稀释比例 | 指标 | 细胞培养上清液 (n=10) | 血清 (n=10) | 肝素血浆 (n=10) | | 1:4 | 预期值均值 (%) | 92
| 92
| 104
| | 1:4 | 范围 (%) | 83-120
| 83-97
| 93-119
| | 1:8 | 预期值均值 (%) | 101
| 97
| 101
| | 1:8 | 范围 (%) | 86-106
| 96-102
| 89-118
| | 1:16 | 预期值均值 (%) | 100
| 90
| 100
| | 1:16 | 范围 (%) | 82-108
| 88-94
| 92-111
| | 1:32 | 预期值均值 (%) | 93
| 98
| 103
| | 1:32 | 范围 (%) | 90-104
| 86-99
| 96-115
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注意事项
1. 浓缩洗涤液低温情况下可能会出现结晶，请水浴加热使结晶完全溶解后再配制工作液；
2. 严禁混用不同批号试剂盒的组分；
3. 加样过程请避免产生气泡，实验操作过程中一定要保证试剂充分混匀，否则会使结果产生较大误差；
4. 说明书中提到的室温条件，请严格控制在25~28℃;
5. 为了您的安全和健康，请穿实验服并戴一次性手套操作；
6. 本产品仅限科研使用。
This ELISA kit adopts the double-antibody sandwich ELISA method to detect the concentration of human CRP in samples. The human CRP capture antibody is pre-coated on the microplate. When samples or standards are added, human CRP binds to the capture antibody, while other unbound free components are removed by washing. Subsequently, biotin-labeled anti-human CRP antibody is added, which binds to human CRP to form a sandwich immune complex; residual free components are then removed by washing. Next, enzyme conjugate is added, and biotin specifically binds to the enzyme conjugate. Thereby, HRP on the enzyme conjugate is linked to the sandwich immune complex, with unbound components removed by another washing step. Finally, chromogenic substrate is added. If human CRP is present in the sample, the bound HRP catalyzes the oxidation of the colorless substrate to produce a blue product. After adding stop solution, the final reaction solution turns yellow. The absorbance value is measured at 450 nm using a microplate reader. The concentration of human CRP is positively correlated with the OD₄₅₀ value. A standard curve is plotted based on standard samples, and the human CRP concentration in unknown samples can be calculated by matching their OD₄₅₀ values against the standard curve.
Background Introduction
C-reactive protein (CRP) is an acute-phase protein whose plasma level rises sharply upon infection or tissue injury in the body. CRP can activate the complement system and enhance the phagocytosis of phagocytes, eliminating invasive pathogenic microorganisms as well as damaged, necrotic and apoptotic tissue cells, thus playing a vital protective role in the innate immune response.
CRP has high affinity for phosphatidylcholine residues and can bind to multiple endogenous and exogenous ligands to activate the classical complement pathway. Meanwhile, it limits the progression and intensity of inflammatory reactions in the late stage of complement activation. In addition, CRP exhibits opsonization and agglutination activities similar to IgG and complement, enhancing the phagocytic capacity of macrophages against various bacteria and foreign substances, and reducing abnormal immune responses caused by exposure to exogenous antigens.
Kit Components
| Item No. | Appearance | Components | 96T | Storage | | H1509775A | — | Pre-coated Assay Plate | 96 wells | 2-8℃. | | H1509775B | Liquid | Sample Diluent | 30 mL | 2-8℃. | | H1509775C | Solid | Recombinant human CRP standard (lyophilized) | 2 vials (50 ng/vial) | 2-8℃. | | H1509775D | Liquid | Biotin-labeled human CRP antibody | 130 μL (Dilution ratio: 1:100) | 2-8℃. | | H1509775E | Liquid | Antibody Diluent | 12 mL | 2-8℃. | | H1509775F | Liquid | Enzyme Conjugate (HRP-labeled Streptavidin) | 130 μL (Dilution ratio: 1:100) | 2-8℃. | | H1509775G | Liquid | Enzyme Conjugate Diluent | 12 mL | 2-8℃. | | H1509775H | Liquid | Concentrated Washing Buffer (25×) | 30 mL | 2-8℃. | | H1509775I | Liquid | TMB Substrate | 10 mL | 2-8℃. | | H1509775J | Liquid | Stop Solution | 10 mL | 2-8℃. | | H1509775K | — | Plate Sealer | 4 sheets | 2-8℃. |
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Operating Procedures
1. Sample Preparation
(1) Select the corresponding pretreatment method according to sample type:
A. Cell Culture Supernatant: Centrifuge the supernatant at 100~500×g for 5 min to remove suspended particles.
B. Serum Sample: Allow whole blood to stand at room temperature for 0.5~2 h for natural coagulation and serum separation. Centrifuge at 4℃, 1000~2000×g for 10 min, collect the yellow supernatant. Avoid aspirating the precipitate. Place prepared serum on ice for standby; do not add preservatives or anticoagulants.
C. Plasma Sample: Anticoagulate whole blood with EDTA, mix well and place on ice. Centrifuge at 4℃, 1000~2000×g for 10 min, collect the yellow supernatant. Avoid aspirating the precipitate and store the prepared plasma on ice.
D. Tissue Homogenate / Body Fluid: Centrifuge to remove precipitates.
Notes:① If detection cannot be performed immediately, aliquot and store samples at -20℃ after preparation; avoid repeated freeze-thaw cycles.
② Ensure samples are clear and transparent. Centrifuge to remove suspended matter before testing if necessary.
③ Do not use hemolytic, icteric, hyperlipidemic or contaminated samples to ensure accurate results.
(2) Sample Dilution
Refer to relevant literature to estimate the content of the target analyte in the sample, so as to determine the appropriate dilution ratio, ensuring the concentration of the target analyte in the diluted sample falls within the optimal detection range of the ELISA kit. Adopt different dilution protocols according to the concentration level of the target analyte:
① When the target analyte concentration ranges from 500 to 5000 ng/mL, dilute the sample at a ratio of 1:100, namely add 3 μL sample into 297 μL sample diluent;
② When the target analyte concentration ranges from 50 to 500 ng/mL, dilute the sample at a ratio of 1:10, namely add 25 μL sample into 225 μL sample diluent;
③ When the target analyte concentration ranges from 0.78 to 50 ng/mL, dilute the sample at a ratio of 1:2, namely add 100 μL sample into 100 μL sample diluent;
④ When the target analyte concentration is ≤ 0.78 ng/mL, the sample generally requires no dilution.
The above protocols are for reference only. Please record the sample dilution method in detail during the experiment.
2. Pre-Assay Preparation
(1) Take the kit out of 4℃ refrigerator and equilibrate at room temperature for 20 min. If stored at -20℃, completely thaw all components first, then equilibrate for 20 min. Store remaining reagents at 4℃ or -20℃ promptly after use.
(2) Dilute the 25× concentrated wash buffer to 1× working wash buffer with double distilled water or deionized water.
(3) Preparation and Dilution of Recombinant Human CRP Standard(Prepare within 2 h before use, operate at room temperature 25~28℃ strictly)
① Prepare 50 ng/mL standard: Add 1 mL sample diluent into the standard vial, let stand for over 15 min, then invert and swirl repeatedly to fully dissolve.
② Perform serial two-fold dilution of the 50 ng/mL standard with sample diluent as follows (maximum concentration: 50 ng/mL; diluent serves as 0 ng/mL blank).
| Tube No. | Diluent Volume (μL) | Reconstituted Standard Volume (μL) | Final Concentration (ng/mL) | | A | 0 | 1000 | 50 | | B | 300 | 300 (from Tube A) | 25 | | C | 300 | 300 (from Tube B) | 12.5 | | D | 300 | 300 (from Tube C) | 6.25 | | E | 300 | 300 (from Tube D) | 3.12 | | F | 300 | 300 (from Tube E) | 1.56 | | G | 300 | 300 (from Tube F) | 0.78 | | H | 300 | 0 | 0 |
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Note: Discard any remaining standard after reconstitution and aliquoting.
(4) Preparation of Biotin-Labeled Anti-Human CRP Antibody Working Solution
① Calculate total volume (prepare an extra 100~200 μL to compensate for operational loss) based on 100 μL per well.
② Prepare working solution at the ratio of 1 μL biotin-labeled antibody to 99 μL antibody diluent, mix gently.
(5) Preparation of Enzyme Conjugate Working Solution (Prepare within 1 h before use)
① Calculate total volume (prepare an extra 100~200 μL for loss) based on 100 μL per well.
② Prepare working solution at the ratio of 1 μL enzyme conjugate to 99 μL enzyme conjugate diluent, mix gently.
3. Detection Protocol
(1) Determine the required number of microplate strips, place them into the plate frame. Seal unused strips in the original aluminum foil bag and store at 4℃ or -20℃.
Notes:① It is recommended to run duplicate wells for standards and samples.
② A standard curve must be generated for each independent assay.
(2) Add 100 μL of diluted samples and serially diluted standards into corresponding wells respectively. Seal the plate with sealing film and incubate at 37℃ for 90 min.
Notes:① If the estimated sample concentration exceeds the upper limit of the standard curve, further dilute the sample before testing.
② Complete the loading process within 10 min to avoid result deviation.
(3) Discard the liquid in the microplate without washing; pat the plate dry on absorbent paper.
(4) Add 100 μL prepared biotin-labeled antibody working solution to each well, seal and incubate at 37℃ for 60 min.
(5) Wash the plate 5 times: add 300 μL 1× wash buffer per well, keep 15~30 s between injection and aspiration. Pat dry on absorbent paper after washing.
Note: Sufficient washing is critical; inadequate washing will cause significant experimental error.
(6) Add 100 μL diluted enzyme conjugate working solution per well, seal and incubate at 37℃ for 30 min in the dark.
(7) Wash the plate 5 times following the same method as step (5).
(8) Add 100 μL TMB substrate per well, seal and react at 37℃ for 10~25 min in the dark.
Notes:① Keep TMB away from oxidants and metals during storage and use.
② Optimal color development time varies with laboratory conditions; obvious blue gradient can be observed in the first 3~4 standard wells when the reaction is sufficient.
(9) Add 100 μL stop solution per well, mix gently. Immediately measure OD₄₅₀ with a microplate reader, set 540 nm or 570 nm as the correction wavelength to obtain corrected absorbance (OD₄₅₀-OD₅₄₀ or OD₄₅₀-OD₅₇₀).
Note: Complete OD value reading within 10 min.
4. Data Analysis
(1) Establish the standard curve: Take standard concentration as the X-axis and OD value as the Y-axis, fit the curve with 4-Parameter Logistic (4-PL) regression using professional software. Calculate sample concentration according to the sample OD value against the standard curve.
Notes:① The assay is valid only when the OD deviation of duplicate wells is within 20%; use the average value as the final reading.
② If the sample OD value exceeds the upper limit of the standard curve, dilute the sample and re-test; multiply the result by the dilution factor when calculating the actual concentration.
(2) Typical Data
This standard curve is provided for demonstration only. A standard curve should be generated for each set of samples assayed.
| Standard concentration (ng/ml) | O.D. | | 0 | 0.087 | | 0.78 | 0.181 | | 1.56 | 0.253 | | 3.12 | 0.457 | | 6.25 | 0.826 | | 12.5 | 1.283 | | 25 | 1.857 | | 50 | 2.316 |
|
(3) Precision
① Intra-Assay Precision (Precision within an assay): Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
② Inter-Assay Precision (Precision between assays): Three samples of known concentration were tested in separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
| Intra-Assay Precision | Intra-Assay Precision | Intra-Assay Precision | Inter-Assay Precision | Inter-Assay Precision | Inter-Assay Precision | | Sample | 1 | 2 | 3 | 1 | 2 | 3 | | n | 20 | 20 | 20 | 20 | 20 | 20 | | Mean (ng/ml) | 64
| 73
| 105
| 84
| 49
| 193
| | Standard Deviation | 5.1
| 2.5
| 5.4
| 2.6
| 1.8
| 8.9
| | CV% | 7.9
| 3.4
| 5.1
| 3.1
| 3.7
| 4.6
|
|
(4) Recovery
The recovery of human C-Reactive Protein/CRP spiked to three levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average Recovery (%) | Range (%) | | Cell culture supernates (n=10) | 113
| 90%-119%
| | Serum (n=10) | 102
| 88%-110%
| | Herparin plasma (n=10) | 97
| 90%-118%
|
|
(5) Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human C-Reactive Protein/CRP were serially diluted with calibrator diluent to produce samples with values within the dynamic range of the assay.
| Dilution Ratio | Index | Cell Culture Supernatants (n=10) | Serum (n=10) | Heparin Plasma (n=10) | | 1:4 | Average of Expected (%) | 92
| 92
| 104
| | 1:4 | Range (%) | 83-120
| 83-97
| 93-119
| | 1:8 | Average of Expected (%) | 101
| 97
| 101
| | 1:8 | Range (%) | 86-106
| 96-102
| 89-118
| | 1:16 | Average of Expected (%) | 100
| 90
| 100
| | 1:16 | Range (%) | 82-108
| 88-94
| 92-111
| | 1:32 | Average of Expected (%) | 93
| 98
| 103
| | 1:32 | Range (%) | 90-104
| 86-99
| 96-115
|
|
Precautions
1. Crystals may form in concentrated washing buffer at low temperature. Warm in water bath to dissolve completely before use.
2. Do not mix components from different kit lots.
3. Avoid bubbles during pipetting. Ensure full mixing to avoid deviation.
4. Room temperature: strictly 25–28 ℃.
5. Wear lab coat and disposable gloves for personal safety.
6. For research use only, not for clinical diagnosis.
ZJ26F0636937
危险品化学品经营许可证（带存储）