
Product Features
Product Name Human MMP-9 EasyTest™ ELISA Kit Assay Type Solid Phase Sandwich ELISA Format 96-well strip plate Assay Process One wash step, two incubations Assay Length 1.5 hours Sample Volume Serum , Plasma, 50µL Detection Method Colorimetric Sensitivity 22pg/mL Assay Range 65.5-16000pg/mL Reactivity Human Cross-Reactivity No cross-reactivity observed with available related molecules Gene Names MMP9; CLG4B; GELB; MANDP2; MMP-9 Gene ID 4318 Swissprot ID P14780
Recovery
Sample Type | Average % Recovery | Range % |
Serum | 90 | 79-106 |
Plasma | 92 | 90-100 |
Linearity
Sample Type | Dilution | % of Expected |
Serum | 1:2 | 118 |
1:4 | 94 | |
1:8 | 88 | |
Plasma | 1:2 | 116 |
1:4 | 98 | |
1:8 | 103 |
Standard Curve: Human MMP-9 EasyTest™ ELISA Kit
Kit Components
• Pre-Coated 96-well Strip Microplate • Wash Buffer • Stop Solution • Assay Diluent • Standards • Bioaim human MMP-9 mix • TMB One-Step Substrate
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the manual. 2. Add 50 µl of standard or sample to each well. 3. Add 50 µl of Bioaim mix. 4. Incubate 1 h at RT. 5. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well. 6. Incubate 15 min at RT. 7. Add 50 µl of Stop Solution to each well. 8. Read at 450 nm/570 nm.
Background
Matrix metallopeptidase 9 (MMP-9), also known as 92 kDa type IV collagenase, 92 kDa gelatinase or gelatinase B (GELB), is a matrixin, a class of enzymes that belong to the zinc-metalloproteinases family involved in the degradation of the extracellular matrix. In humans the MMP9 gene. Proteins of the matrix metalloproteinase (MMP) family are involved in the breakdown of extracellular matrix in normal physiological processes, such as embryonic development, reproduction, angiogenesis, bone development, wound healing, cell migration, learning and memory, as well as in pathological processes, such as arthritis, intracerebral hemorrhage, and metastasis.
Studies in rhesus monkeys suggest that the enzyme is involved in IL-8-induced mobilization of hematopoietic progenitor cells from bone marrow, and murine studies suggest a role in tumor-associated tissue remodeling. Thrombospondins, intervertebral disc proteins, regulate interaction with matrix metalloproteinases (MMPs) 2 and 9, which are key effectors of ECM remodeling.
Storage
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Regulatory Status
Human MMP-9 EasyTest™ ELISA Kit is for Research Use Only.
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的试剂:免疫吸附剂、结合物和酶的底物。
完整的ELISA试剂盒包含以下各组分:
(1)已包被抗原或抗体的固相载体(免疫吸附剂);
(2)酶标记的抗原或抗体(结合物);
(3)酶的底物;
(4)阴性对照品和阳性对照品(定性测定中),参考标准品和控制血清(定量测定
中);
(5)结合物及标本的稀释液;
(6)洗涤液;
(7)酶反应终止液。
今天文章就简单整理了一下最近遇到的问题,有些问题对你来说可能很简单,但是还是会给一些科研菜鸟带去小麻烦,特整理了这么个专场来涨涨姿势。
这里就简单整理了5个问题,具体内容如下:
小编自己在各大论坛看到最多的三个答案:
1)48T可做42个样本加6个标准曲线;96T可做90个样本加6个标准曲线
2)以96T试剂盒来算,定量的试剂盒一般可以做90个左右的样品,定性的试剂盒可以做94个左右的样品
3)96T的ELISA试剂盒,去除标准品复孔检测,最多还能检测80个标本;48T的最多30个样本。
2、elisa试剂盒96T的能做检测多少血清样品?板能重复利用吗?
96T,一般做复孔的话,做标准曲线需要16个孔,剩下80个孔就可以做40个样本。这个板不能重复利用。而且配套的试剂也只是够一块板子用的。
3、想请教一下,我想用elisa法测血清里的一种抗原,查了资料,发现没有现成的试剂盒用,现在就有2种备选方案,一种是买抗体对自己包被(查了R&D等大公司都没有可用于elisa的单抗)。哪种比较好呢?自己包被的会不会比国产的试剂盒可靠呢?
如果你觉得技术不错有队其他公司不放心,可以自己购买抗体对自己包被,这个就是麻烦点,自己包被用着放心,舒心;
另外国内还是有一些资质不错的公司,如北京达科为,上海巧伊,依科赛,深圳欣博盛等等
4、western和ELISA处理样本有何差别?
WesternBlot样品制备要加蛋白裂解液(含有蛋白酶抑制剂),然后冰上研磨,收集于EP管,冰上裂解30min,4度12000rpm离心15min,取上清分装,即得到蛋白样品。建议直接电泳,多余的-80保存。建议分装的蛋白每支略多于一次上样量,一次性用完一支,避免反复冻融。
ELISA样品,直接脑组织冰上匀浆,收集于EP管,同上一样离心,大约220-250ul每支分装(因为ELISA上样量基本都是每孔100ul,做复孔,每个样品2孔,就需要200ul,为了避免不够2孔,分装时要大于200ul每支),直接实验。多余的-80保存,一样避免反复冻融。
不过ELISA最好少量样品进行预实验,或者查阅文献,看看大概组织里有多高浓度,选择合适的试剂盒,测量范围要与组织浓度相符。避免浓度过高测量不准,或者浓度过低,小于试剂盒最低可测浓度。
5、Elisa试剂盒该买哪家公司的呢?
提出这个问题的朋友其实该打,目前国际上各个品牌的试剂盒生厂商,都会有其偏重点,像R&D在细胞因子方面的试剂盒就非常牛逼,而其他牌子也一样的。
Bender:粘附分子方面的试剂盒;
Cayman:花生四烯酸类产品的试剂盒;
Merck(Linco):内分泌研究方面的试剂盒;
Tips:
1、买Elisa试剂盒时,尽量先查文献,通过文献找品牌,再找供应商,找供应商就找正规代理商,千万不要去干“赔夫人又折兵”的蠢事。
2、ELISA中,复孔就是同一个样品加到至少两个孔中(一般3个或4个),最后取其平均值作为样品的读数,这样测得的结果才有说服力!
做复孔是为了较好的较少组内差异,如果血清不足不做复孔也可以。