Omni-Rapid™快速蛋白定量试剂盒
Omni-Rapid™Protein Assay Kit
本产品冰袋运输; 即用型BSA标准品 -20℃保存,其它组分4℃保存,保质期12个月。
货号规格
goodZJ103 500次(微孔)
产品内容
组分 |
体积 |
试剂A |
100 mL |
试剂B |
3 mL |
即用型BSA标准品①( 0 μg/mL ) |
1 mL |
即用型BSA标准品②(125 μg/mL) |
1 mL |
即用型BSA标准品③(250 μg/mL) |
1 mL |
即用型BSA标准品④(500 μg/mL) |
1 mL |
即用型BSA标准品⑤(750 μg/mL) |
1 mL |
即用型BSA标准品⑥(1000 μg/mL) |
1 mL |
即用型BSA标准品⑦(1500 μg/mL) |
1 mL |
即用型BSA标准品⑧(2000 μg/mL) |
1 mL |
产品特点
g
d简单快速——室温5 min完成显色反应;
g
d方便快捷——提供即用型标准品,省去繁琐的稀释步骤;
g
d准确性高——变异系数远小于考马斯亮蓝染色法;
g
d线性范围宽——灵敏,检测范围:20~2000 μg/mL;
g
d兼容性好——与大多数金属离子、螯合剂及去污剂兼容性较好。
产品简介
goodOmni-Rapid™快速蛋白定量试剂盒的原理与传统BCA蛋白定量法类似,但采用了一种不同于BCA(Bicin-choninic Acid)的全新特殊的螯合剂,从而实现了对蛋白质浓度进行快速、稳定、灵敏的测定。其原理是在碱性环境下蛋白质分子中的肽键能与Cu2+形成络合物,将Cu2+还原成Cu+,Cu+与螯合物结合,从而发生颜色反应。本试剂盒中的螯合剂可敏感特异地与Cu+结合,只需室温孵育5 min即可形成稳定的橙黄色水溶性复合物,而传统BCA法则需在37℃下孵育30 min才可完成颜色反应。该橙黄色的复合物在480 nm处有强光吸收值,颜色的深浅与蛋白质浓度成正比,可根据吸收值的大小来测定蛋白质的含量。本试剂盒含有一系列浓度的蛋白质标准品溶液(BSA溶液),即取即用,无需稀释,方便快捷。
使用说明
good◈以微孔酶标仪法为例:
gooddd1. 配置显色工作液:
gooddd1. a. 计算显色工作液总量:
goodgoodgoodd工作液总量 = (BSA标准品样本个数+待测样本个数)×复孔数×每个样本显色工作液体积
gooddd1. a. 举例:BSA标准品样本个数为8个,待测样本个数3个,复孔数3个。
goodgoodgoodd显色工作液总量 = (8个BSA标准品样本+3个待测样本)×3个复孔×200 μL(每个样本工作液体积) = 6.6 mL
gooddd1. b. 根据计算出的显色工作液需要总量,将试剂A和试剂B按照50:1的体积比,配制显色工作液,充分混匀。
gooddd1. b. 注意:1) 试剂B刚加入试剂A时,会出现灰蓝色沉淀,但只需混匀几秒钟,沉淀就会消失,形成透亮的绿色溶液;
gooddd1. b. 注意:2)建议工作液现用现配,在室温下,工作液会逐渐变为深绿色,但只要在1.5 h内使用,对定量的准确性不会造成影响;
gooddd1. b. 注意:3) 由于加样可能存在误差,建议配制BCA工作液时,多配制1~2个孔。
gooddd2. 定量检测
gooddd3. (1)分别取 即用型BSA标准品①~⑧ 各20 μL加到96孔板中( BSA标准品 使用前须充分溶解摇匀);
孔号 |
1 |
2 |
3 |
4 |
5 |
6 |
7 |
8 |
添加物 |
标准品① |
标准品② |
标准品③ |
标准品④ |
标准品⑤ |
标准品⑥ |
标准品⑦ |
标准品⑧ |
体积(μL) |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
20 |
BSA终浓度(μg/mL) |
0 |
125 |
250 |
500 |
750 |
1000 |
1500 |
2000 |
gooddd3. (2)用1×PBS或0.9%生理盐水将样品适当稀释(可以多作几个梯度,如2倍、4倍、8倍稀释),加20 μL到96孔板的样品孔中;
gooddd3. (3)各孔加入200 μL显色工作液,充分混匀,盖上96孔板盖,室温孵育5min,即可进行检测;
gooddd3. (3)注意:由于颜色反应速度较快,须保证在20~30 min之内完成读值。如果必须在30 min后才能读值,可提前加入50 μL 1 M HCl终止反应。
gooddd3. (4)用酶标仪测定每个样品及BSA标准品的A480,注意要减去空白对照( 标准品① +工作液)的A480;
gooddd3. (5)绘制标准曲线,计算样品中的蛋白浓度。
gooddd3. (3)注意:数据处理时需要去除明显错误的值。待测样品浓度可以从标准曲线中查得, 实际浓度需要乘以样品的稀释倍数。如果是计算机绘制的曲线,可以从计算机给出的线性方程式计算出待测样品的浓度。
注意事项
good1. 本产品可以采用酶标仪(微孔检测法)或者分光光度计(试管检测法)测定蛋白浓度,如使用普通的分光光度计测定,需根据比色皿的最小检测体积,适当加大BCA工作液的用量使不小于最小检测体积,样品和标准品的用量可相应按比例放大也可不变。使用分光光度计测定蛋白浓度时,每个试剂盒可以测定的样品数量可能会显著减少;
good2. 建议每次测定蛋白样品时,都须绘制标准曲线,以获得准确数据;
good3. 完成室温孵育5 min后,须在20~30 min内完成检测,否则会影响蛋白定量的准确度;
good4. 如待测样品中含较多的干扰物质(具体见附表),可采用其它蛋白定量产品;
good5. 为了您的安全和健康,请穿实验服并戴一次性手套操作;
good6. 本产品仅限科研使用。
干扰物质附表
化合物 |
耐受浓度 |
化合物 |
耐受浓度 |
盐/缓冲液 |
去垢剂 |
ACES, pH 7.8 |
25mM |
Brij-35 |
5.0% |
Bicine, pH 8.4 |
20mM |
Brij-58 |
1.0% |
Borate |
50mM |
Na Deoxycholate(DOC) |
5.0% |
Calcium chloride in TBS, pH 7.2 |
10mM |
Octyl β-glucoside |
5.0% |
Na-Carbonate/Na-Bicarbonate, pH 9.4 |
0.1M |
Span-20 |
0.5% |
Cesium bicarbonate |
100mM |
TritonX-100 |
5.0% |
CHES, pH 9.0 |
100mM |
Triton-X-114, riton-X-305,Triton-X-405 |
1.0% |
Na-Citrate |
75mM |
Tween-20, Tween-60, Tween-80 |
5.0% |
MOPS, pH 7.2 |
100mM |
CHAPS |
5.0% |
Cobalt chloride in TBS, pH 7.2 |
0.8mM |
CHAPSO |
5.0% |
EPPS, pH 8.0 |
100mM |
Zwittergent 3-14 |
1.0% |
Ferric chloride in TBS, pH 7.2 |
10mM |
螯合剂 |
Glycine•HCl, pH 2.8 |
100mM |
EDTA |
10mM |
Ammonium sulfate |
Ø |
Sodium citrate |
200mM |
Guanidine•HCl |
4M |
还原剂 |
HEPES, pH 7.5 |
100mM |
2-mercaptoethanol |
Ø |
Imidazole, pH 7.0 |
12.5mM |
Dithiothreitol (DTT) |
Ø |
MES, pH 6.1 |
100mM |
N-acetylglucosamine in PBS, pH 7.2 |
10mM |
MES (0.1M), NaCl (0.9%), pH 4.7 |
Undiluted |
糖类 |
Nickel chloride in TBS, pH 7.2 |
10mM |
Glucose |
10mM |
PBS; Phosphate (0.1 M), NaCl (0.15 M), pH 7.2 |
Undiluted |
其它 |
A-cetone |
10% |
PIPES, pH 6.8 |
100mM |
Acetonitrile |
10% |
RIPA lysis buffer: 50 mM Tris, 150 mM NaCl, 0.5% DOC, 1% NP-40, 0.1% SDS, pH 8.0 |
Undiluted |
Aprotinin |
10mg/L |
DMF |
10% |
DMSO |
10% |
Sodium acetate, pH 4.8 |
200 mM |
Ethanol |
10% |
Sodium azide |
0.2% |
Glycerol (fresh) |
10% |
Sodium bicarbonate |
100mM |
Hydrochloric acid |
100mM |
Sodium chloride |
1 M |
Leupeptin |
10mg/L |
Sodium citrate, pH 4.8 or pH 6.4 |
200mM |
Methanol |
10% |
Sodium phosphate |
100mM |
N/A |
Tricine, pH 8.0 |
25mM |
Triethanolamine, pH 7.8 |
25mM |
Tris |
250mM |
TBS: Tris (25 mM), NaCl (0.15 M), pH 7.6 |
Undiluted |
Tris (25 mM), Glycine (192 mM), pH 8.0 |
1:2 dilution |
相关论文
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