人*脱羧酶65抗体（GADA65-Ab）ELISA检测试剂盒
检测原理
试剂盒采用间接法酶联免疫吸附试验（elisa）。往预先包被人*脱羧酶65抗体（gada65-ab）捕获抗原的包被微孔中，依次加入标本、标准品、hrp标记的检测抗体，经过温育并*洗涤。用底物tmb显色，tmb在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成终的黄色。颜色的深浅和样品中的人*脱羧酶65抗体（gada65-ab）呈正相关。用酶标仪在450nm 波长下测定吸光度（od 值），计算样品浓度。
样品收集、处理及保存方法
1. 血清：使用不含热原和内毒素的试管，操作过程中避免任何细胞刺激，收集血液后，3000转离心10分钟将血清和红细胞迅速小心地分离。
2. 血浆：edta、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。
3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。
4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。
5. 保存：如果样本收集后不及时检测，请按一次用量分装，冻存于-20℃，避免反复冻融，在室温下解冻并确保样品均匀地充分解冻。
自备物品
酶标仪（450nm）高精度加样器及枪头：0.5-10ul、2-20ul、20-200ul、200-1000ul37℃恒温箱操作注意事项
试剂盒保存在2-8℃，使用前室温平衡20分钟。从冰箱取出的浓缩洗涤液会有结晶，这属于正常现象，水浴加热使结晶*溶解后再使用。实验中不用的板条应立即放回自封袋中，密封（低温干燥）保存。预处理后的样本无需稀释，直接取10μl加样即可。严格按照说明书中标明的时间、加液量及顺序进行温育操作。所有液体组分使用前充分摇匀。试剂盒组成
名称
96孔配置
48孔配置
备注
微孔酶标板
12孔×8条
12孔×4条
无
标准品
0.3ml
0.3ml
无
*
6ml
3ml
无
检测抗体-hrp
10ml
5ml
无
20×洗涤缓冲液
25ml
15ml
按说明书进行稀释
底物a
6ml
3ml
无
底物b
6ml
3ml
无
终止液
6ml
3ml
无
封板膜
2张
2张
无
说明书
1份
1份
无
自封袋
1个
1个
无
注：标准品浓度依次为：800、400、200、100、50、0 ng/ml.
试剂的准备
20×洗涤缓冲液的稀释：蒸馏水按1：20稀释，即1份的20×洗涤缓冲液加19份的蒸馏水。
洗板方法
手工洗板：甩尽孔内液体，每孔加满洗涤液，静置1min后甩尽孔内液体，在吸水纸上拍干，如此洗板5次。自动洗板机：每孔注入洗液350μl，浸泡1min，洗板5次。操作步骤
从室温平衡20min后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回4℃。设置标准品孔和样本孔，标准品孔各加不同浓度的标准品50μl；待测样本孔先加待测样本10μl，再加*40μl；随后标准品孔和样本孔中每孔加入辣根过氧化物酶（hrp）标记的检测抗体100μl，用封板膜封住反应孔，37℃水浴锅或恒温箱温育60min。弃去液体，吸水纸上拍干，每孔加满洗涤液，静置1min，甩去洗涤液，吸水纸上拍干，如此重复洗板5次（也可用洗板机洗板）。每孔加入底物a、b各50μl，37℃避光孵育15min。每孔加入终止液50μl，15min内，在450nm波长处测定各孔的od值。结果判断
绘制标准曲线：在excel工作表中，以标准品浓度作横坐标，对应od值作纵坐标，绘制出标准品线性回归曲线，按曲线方程计算各样本浓度值。
试剂盒性能
准确性：标准品线性回归与预期浓度相关系数r值，大于等于0.9900。灵敏度：低检测浓度小于1.0ng/ml。特异性：不与其它可溶性结构类似物交叉反应。重复性：板内变异系数小于10%、板间变异系数小于15%。贮藏：2-8℃，避光防潮保存。有效期：6个月免责声明
试剂盒仅供研究使用，不得用于临床实验或人体实验，否则所产生的一切后果，由实验者承担，本公司概不负责。严格按照说明书操作，实验者违反说明书操作，后果由实验者承担。
for research use only.
not for use in diagnostic procedures.
humanglutamate decarboxylase 65 antibody(gada65-ab) elisa kit instruction
intended use
this gada65-abelisa kit is intended laboratory for research use only and is not for use in diagnostic or therapeutic procedures.the stop solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. in order to measure the concentration of gada65-abin the sample, this gada65-abelisa kit includes a set of calibration standards. the calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of optical density versus gada65-abconcentration. the concentration of gada65-abin the samples is then determined by comparing the o.d. of the samples to the standard curve.
samplecollection and storages
serum- use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.avoid repeated freeze-thaw cycles
plasma- collect plasma using edta or heparin as an anticoagulant. centrifuge samples for 30minutes at 3000×g at 2-8℃ within 30 minutes of collection. store samples at -20℃or -80℃. avoid repeated freeze-thaw cycles.
cell culture supernates and other biological fluids-remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. avoid repeated freeze-thaw cycles.
note: the samples shouldbe centrifugated adequately and no hemolysis or granule was allowed.
materials required but not supplied
1. standard microplate reader(450nm)
2. precision pipettes and disposable pipette tips.
3. 37 ℃ incubator
precautions
1. donotsubstitutereagentsfromone kitto another.standard, conjugateandmicroplates are matchedfor optimal performance.useonly thereagentssuppliedby manufacturer.
2. donotremovemicroplatefromthe storage baguntilneeded.unusedstripsshouldbe stored at2-8°cin their pouchwiththe desiccantprovided.
3. mix all reagents before using.
remove allkitreagentsfromrefrigerator and allowthemto reachroomtemperature( 20-25°c)
materials supplied
name
96determinations
48determinations
microelisa stripplate
12*8strips
12*4strips
standard
0.3ml
0.3ml
sample diluent
6.0ml
3.0ml
hrp-conjugate reagent
10.0ml
5.0ml
20x wash solution
25ml
15ml
chromogen solution a
6.0ml
3.0ml
chromogen solution b
6.0ml
3.0ml
stop solution
6.0ml
3.0ml
closure plate membrane
2
2
user manual
1
1
sealed bags
1
1
note: standard concentration was followed by:
800、400、200、100、50、0 ng/ml.
reagent preparation
20×wash solution:dilute with distilled or deionized water1:20.
assay procedure
1. prepare allreagentsbeforestartingassayprocedure.itisrecommendedthatallstandardsand samplesbe addedin duplicateto the microelisastripplate.
2. add standard: set standard wells, testing sample wells. add standard 50μl to standard well.
3. add sample: add testing sample10μl then add sample diluent 40μl to testing sample well; blank welldoesn’t add anyting.
4. add100μlofhrp-conjugate reagentto each well,cover with anadhesive stripandincubatefor60 minutesat37°c.
5. aspirate each well and wash, repeating the process fourtimes for a total of five washes.wash by filling each well with wash solution(400μl) using a squirt bottle, manifolddispenseror autowasher. complete removal of liquid at each step is essential to good performance. after the last wash, remove any remaining wash solutionby aspirating ordecanting. invert the plate and blot it against clean paper towels.
6. add chromogen solution a 50μl and chromogen solution b 50μl to each well.gently mix and incubate for 15 minutes at 37°c. protect from light.
7. add 50μl stop solution to each well. the color in the wells should change from blue toyellow. if the color in the wells is green or the color change does not
appear uniform,gently tap the plate to ensure thorough mixing.
8. readtheopticaldensity(o.d.)at450nmusinga microtiterplatereaderwithin15minutes.
calculation of results
this standard curve is used to determine the amount in an unknown sample. the standard curve is generated by plotting the average o.d. (450 nm) obtained for each of the six standard concentrations on the vertical (y) axis versus the corresponding concentration on the horizontal (x) axis. first, calculate the mean o.d. value for each standard and sample. all o.d. values, are subtracted by the mean value of the zero standard before result interpretation. construct the standard curve using graph paper or statistical software. to determine the amount in each sample, first locate the o.d. value on the y-axis and extend a horizontal line to the standard curve. at the point of intersection, draw a vertical line to the x-axis and read the corresponding concentration. any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. each user should obtain their own standard curve.the sensitivity by this assay is1.0 ng/ml.standard curve
storage： 2-8℃.
validity： six months.
for research use only; not for therapeutic or diagnostic applications!please read through entire procedure before beginning!
机械隔膜计量泵控制方法的*
EDI超纯水设备推动电子产业信息化进程
中走丝电火花线切割在动模、定模和工具电极加工中的应用
MN1H-2-1/2-MS费斯托电磁阀
注射级山茶油特点2020药典备案
人*脱羧酶65抗体（GADA65-Ab）ELISA检测试剂盒
有机合成危险操作、失误操作及防护（十二-安全汞操作）
EUCHNER安士能开关的特点
意大利ATOS流量阀通过恒定各个支路的流量来实现整个系统流量的均衡输配
通用压缩空气露点仪在设计时需要考虑这些因素
步入大数据时代：泛海三江携手华为 共商IoT生态发展
数控设备管理与维修实例分析
AVTRON编码器
80立方/天一体化生活污水处理设备
复合电吸附装置工作原理和技术参数
GX12CCB直流接触器GIGAVAC继电器上海销售中心
油田污水专用聚丙烯酰胺PAM絮凝剂的使用剂量
聚碳酸酯通用工程塑料拉力机HT140PT
蒸汽疏水阀的维护与保养要点
高温齿轮润滑脂，金属齿轮润滑脂，特种齿轮润滑脂
试剂盒采用间接法酶联免疫吸附试验（elisa）。往预先包被人*脱羧酶65抗体（gada65-ab）捕获抗原的包被微孔中，依次加入标本、标准品、hrp标记的检测抗体，经过温育并*洗涤。用底物tmb显色，tmb在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成终的黄色。颜色的深浅和样品中的人*脱羧酶65抗体（gada65-ab）呈正相关。用酶标仪在450nm 波长下测定吸光度（od 值），计算样品浓度。
样品收集、处理及保存方法
1. 血清：使用不含热原和内毒素的试管，操作过程中避免任何细胞刺激，收集血液后，3000转离心10分钟将血清和红细胞迅速小心地分离。
2. 血浆：edta、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。
3. 细胞上清液：3000转离心10分钟去除颗粒和聚合物。
4. 组织匀浆：将组织加入适量生理盐水捣碎。3000转离心10分钟取上清。
5. 保存：如果样本收集后不及时检测，请按一次用量分装，冻存于-20℃，避免反复冻融，在室温下解冻并确保样品均匀地充分解冻。
自备物品
酶标仪（450nm）高精度加样器及枪头：0.5-10ul、2-20ul、20-200ul、200-1000ul37℃恒温箱操作注意事项
试剂盒保存在2-8℃，使用前室温平衡20分钟。从冰箱取出的浓缩洗涤液会有结晶，这属于正常现象，水浴加热使结晶*溶解后再使用。实验中不用的板条应立即放回自封袋中，密封（低温干燥）保存。预处理后的样本无需稀释，直接取10μl加样即可。严格按照说明书中标明的时间、加液量及顺序进行温育操作。所有液体组分使用前充分摇匀。试剂盒组成
名称
96孔配置
48孔配置
备注
微孔酶标板
12孔×8条
12孔×4条
无
标准品
0.3ml
0.3ml
无
*
6ml
3ml
无
检测抗体-hrp
10ml
5ml
无
20×洗涤缓冲液
25ml
15ml
按说明书进行稀释
底物a
6ml
3ml
无
底物b
6ml
3ml
无
终止液
6ml
3ml
无
封板膜
2张
2张
无
说明书
1份
1份
无
自封袋
1个
1个
无
注：标准品浓度依次为：800、400、200、100、50、0 ng/ml.
试剂的准备
20×洗涤缓冲液的稀释：蒸馏水按1：20稀释，即1份的20×洗涤缓冲液加19份的蒸馏水。
洗板方法
手工洗板：甩尽孔内液体，每孔加满洗涤液，静置1min后甩尽孔内液体，在吸水纸上拍干，如此洗板5次。自动洗板机：每孔注入洗液350μl，浸泡1min，洗板5次。操作步骤
从室温平衡20min后的铝箔袋中取出所需板条，剩余板条用自封袋密封放回4℃。设置标准品孔和样本孔，标准品孔各加不同浓度的标准品50μl；待测样本孔先加待测样本10μl，再加*40μl；随后标准品孔和样本孔中每孔加入辣根过氧化物酶（hrp）标记的检测抗体100μl，用封板膜封住反应孔，37℃水浴锅或恒温箱温育60min。弃去液体，吸水纸上拍干，每孔加满洗涤液，静置1min，甩去洗涤液，吸水纸上拍干，如此重复洗板5次（也可用洗板机洗板）。每孔加入底物a、b各50μl，37℃避光孵育15min。每孔加入终止液50μl，15min内，在450nm波长处测定各孔的od值。结果判断
绘制标准曲线：在excel工作表中，以标准品浓度作横坐标，对应od值作纵坐标，绘制出标准品线性回归曲线，按曲线方程计算各样本浓度值。
试剂盒性能
准确性：标准品线性回归与预期浓度相关系数r值，大于等于0.9900。灵敏度：低检测浓度小于1.0ng/ml。特异性：不与其它可溶性结构类似物交叉反应。重复性：板内变异系数小于10%、板间变异系数小于15%。贮藏：2-8℃，避光防潮保存。有效期：6个月免责声明
试剂盒仅供研究使用，不得用于临床实验或人体实验，否则所产生的一切后果，由实验者承担，本公司概不负责。严格按照说明书操作，实验者违反说明书操作，后果由实验者承担。
for research use only.
not for use in diagnostic procedures.
humanglutamate decarboxylase 65 antibody(gada65-ab) elisa kit instruction
intended use
this gada65-abelisa kit is intended laboratory for research use only and is not for use in diagnostic or therapeutic procedures.the stop solution changes the color from blue to yellow and the intensity of the color is measured at 450 nm using a spectrophotometer. in order to measure the concentration of gada65-abin the sample, this gada65-abelisa kit includes a set of calibration standards. the calibration standards are assayed at the same time as the samples and allow the operator to produce a standard curve of optical density versus gada65-abconcentration. the concentration of gada65-abin the samples is then determined by comparing the o.d. of the samples to the standard curve.
samplecollection and storages
serum- use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. remove serum and assay immediately or aliquot and store samples at -20℃ or -80℃.avoid repeated freeze-thaw cycles
plasma- collect plasma using edta or heparin as an anticoagulant. centrifuge samples for 30minutes at 3000×g at 2-8℃ within 30 minutes of collection. store samples at -20℃or -80℃. avoid repeated freeze-thaw cycles.
cell culture supernates and other biological fluids-remove particulates by centrifugation and assay immediately or aliquot and store samples at -20℃or -80℃. avoid repeated freeze-thaw cycles.
note: the samples shouldbe centrifugated adequately and no hemolysis or granule was allowed.
materials required but not supplied
1. standard microplate reader(450nm)
2. precision pipettes and disposable pipette tips.
3. 37 ℃ incubator
precautions
1. donotsubstitutereagentsfromone kitto another.standard, conjugateandmicroplates are matchedfor optimal performance.useonly thereagentssuppliedby manufacturer.
2. donotremovemicroplatefromthe storage baguntilneeded.unusedstripsshouldbe stored at2-8°cin their pouchwiththe desiccantprovided.
3. mix all reagents before using.
remove allkitreagentsfromrefrigerator and allowthemto reachroomtemperature( 20-25°c)
materials supplied
name
96determinations
48determinations
microelisa stripplate
12*8strips
12*4strips
standard
0.3ml
0.3ml
sample diluent
6.0ml
3.0ml
hrp-conjugate reagent
10.0ml
5.0ml
20x wash solution
25ml
15ml
chromogen solution a
6.0ml
3.0ml
chromogen solution b
6.0ml
3.0ml
stop solution
6.0ml
3.0ml
closure plate membrane
2
2
user manual
1
1
sealed bags
1
1
note: standard concentration was followed by:
800、400、200、100、50、0 ng/ml.
reagent preparation
20×wash solution:dilute with distilled or deionized water1:20.
assay procedure
1. prepare allreagentsbeforestartingassayprocedure.itisrecommendedthatallstandardsand samplesbe addedin duplicateto the microelisastripplate.
2. add standard: set standard wells, testing sample wells. add standard 50μl to standard well.
3. add sample: add testing sample10μl then add sample diluent 40μl to testing sample well; blank welldoesn’t add anyting.
4. add100μlofhrp-conjugate reagentto each well,cover with anadhesive stripandincubatefor60 minutesat37°c.
5. aspirate each well and wash, repeating the process fourtimes for a total of five washes.wash by filling each well with wash solution(400μl) using a squirt bottle, manifolddispenseror autowasher. complete removal of liquid at each step is essential to good performance. after the last wash, remove any remaining wash solutionby aspirating ordecanting. invert the plate and blot it against clean paper towels.
6. add chromogen solution a 50μl and chromogen solution b 50μl to each well.gently mix and incubate for 15 minutes at 37°c. protect from light.
7. add 50μl stop solution to each well. the color in the wells should change from blue toyellow. if the color in the wells is green or the color change does not
appear uniform,gently tap the plate to ensure thorough mixing.
8. readtheopticaldensity(o.d.)at450nmusinga microtiterplatereaderwithin15minutes.
calculation of results
this standard curve is used to determine the amount in an unknown sample. the standard curve is generated by plotting the average o.d. (450 nm) obtained for each of the six standard concentrations on the vertical (y) axis versus the corresponding concentration on the horizontal (x) axis. first, calculate the mean o.d. value for each standard and sample. all o.d. values, are subtracted by the mean value of the zero standard before result interpretation. construct the standard curve using graph paper or statistical software. to determine the amount in each sample, first locate the o.d. value on the y-axis and extend a horizontal line to the standard curve. at the point of intersection, draw a vertical line to the x-axis and read the corresponding concentration. any variation in operator, pipetting and washing technique, incubation time or temperature, and kit age can cause variation in result. each user should obtain their own standard curve.the sensitivity by this assay is1.0 ng/ml.standard curve
storage： 2-8℃.
validity： six months.
for research use only; not for therapeutic or diagnostic applications!please read through entire procedure before beginning!
机械隔膜计量泵控制方法的*
EDI超纯水设备推动电子产业信息化进程
中走丝电火花线切割在动模、定模和工具电极加工中的应用
MN1H-2-1/2-MS费斯托电磁阀
注射级山茶油特点2020药典备案
人*脱羧酶65抗体（GADA65-Ab）ELISA检测试剂盒
有机合成危险操作、失误操作及防护（十二-安全汞操作）
EUCHNER安士能开关的特点
意大利ATOS流量阀通过恒定各个支路的流量来实现整个系统流量的均衡输配
通用压缩空气露点仪在设计时需要考虑这些因素
步入大数据时代：泛海三江携手华为 共商IoT生态发展
数控设备管理与维修实例分析
AVTRON编码器
80立方/天一体化生活污水处理设备
复合电吸附装置工作原理和技术参数
GX12CCB直流接触器GIGAVAC继电器上海销售中心
油田污水专用聚丙烯酰胺PAM絮凝剂的使用剂量
聚碳酸酯通用工程塑料拉力机HT140PT
蒸汽疏水阀的维护与保养要点
高温齿轮润滑脂，金属齿轮润滑脂，特种齿轮润滑脂