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應用
用适当的过氧化物酶底物检测含有α-D-甘露糖、α-D-葡萄糖的糖蛋白
通用Western Blot实验方法:
- 糖蛋白样品规格:500ng
- 凝集素浓度:0.1ug/ml
- 每条泳道上样500 ng糖蛋白样品
- 进行4-20% Bis-Tris SDS page凝胶
- 转膜至PVDF膜上
- 用于RIPA缓冲液(R0278 Sigma)在室温对膜进行封闭1小时
- 在室温用RIPA缓冲液以0.1 ug/ml孵育HRP凝集素2小时
- 用25ml RIPA缓冲液洗膜 5 x 5分钟
- 用化学发光底物(CPS1-120)进行检测
生化/生理作用
刀豆素 A 无血型特异性,但对末端 α-D-甘露糖基和 α-D-葡糖基残基具有亲和力。需要 Ca2+ 和 Mn2+ 离子存在才能显示活性。刀豆素 A 在 pH 6.5 或更低的条件下解离为二聚体。pH 在 5.8 至 7.0 之间时,刀豆素 A 以四聚体形式存在;pH 高于 7.0 时,形成更高的聚合体。根据聚合程度不同,刀豆素 A 可能显示出具有促有丝分裂的活性。琥珀酰化可以生成一种能够在 pH 高于 5.6 时仍保持二聚体形式的活性二聚体。
包裝
包装规格取决于蛋白质含量
單位定義
一单位在20℃、pH 6.0条件下,在20秒内从邻苯三酚形成1 mg的红棓酚。
外觀
含有柠檬酸三钠缓冲盐和微量钙和锰的冻干粉末
準備報告
使用Wilson和Nakane的方法从VI型过氧化物酶(P8375)制备而得,其可促进耦合但能防止Con A和过氧化物酶糖残基之间的相互作用。
分析報告
在报道中,凝集活性以μg/ml表示,并且是在含有1mg/mL Ca 2+和Mn 2+的磷酸盐缓冲盐水(pH 6.8)的系列稀释液中进行测定的。该活性是在25℃孵育1小时后,对2%人红细胞悬浮液进行凝集所需的最低浓度。
訊號詞
Danger
危險聲明
危險分類
Repr. 2 - Resp. Sens. 1 - Skin Sens. 1
儲存類別代碼
11 - Combustible Solids
水污染物質分類(WGK)
WGK 3
閃點(°F)
Not applicable
閃點(°C)
Not applicable
個人防護裝備
Eyeshields, Faceshields, Gloves, type P3 (EN 143) respirator cartridges
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