Format : | Purified |
Amount : | 100 µg |
Isotype : | Rabbit IgG |
Purification : | Protein A Chromatography |
Content : | 25 µg in 50 µl/100 µg in 200 µl PBS containing 0.05% BSA and 0.05% sodium azide. Sodium azide is highly toxic. |
Storage condition : | Store the antibody at 4°C, stable for 6 months. For long-term storage, store at -20°C. Avoid repeated freeze and thaw cycles. |
Gene : | GAPDH |
Gene ID : | 2597 |
Uniprot ID : | P04406 |
Alternative Name : | GAPDH, GAPD, CDABP0047, OK/SW-cl.12 |
Immunogen Information : | A partial length recombinant GAPDH protein was used as the immunogen for this antibody. |
GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) is an enzyme best known for its role in glycolysis. However, extra-glycolytic functions of GAPDH have been described, including regulation of protein expression via RNA binding. GAPDH binds to numerous AREs (adenine-uridine rich elements) from various mRNA 3'-untranslated regions in vitro and in vivo despite its lack of a canonical RNA binding motif. GAPDH specifically catalyzes the simultaneous phosphorylation and oxidation of glyceraldehyde 3-phosphate using NAD+ (Nicotinamide Adenine Dinucleotide) as a cofactor to produce glycerate 1,3-biphosphate and NADH. In addition to its role in energy production, GAPDH has been implicated in many cellular processes including DNA repair tRNA export, membrane fusion and transport, endocytosis and nuclear membrane assembly, and cell death.
Western blot analysis: 1-2 µg/ml
For Research Use Only. Not for use in diagnostic/therapeutics procedures.
Subcellular location: | Cytoplasm, Nucleus, Cytoplasm, Membrane, Cytoplasm |
Post transnational modification: | Oxidative stress can promote the formation of high molecular weight disulfide-linked GAPDH aggregates, through a process called nucleocytoplasmic coagulation. Such aggregates can be observed in vivo in the affected tissues of patients with Alzheimer disease or alcoholic liver cirrhosis, or in cell cultures during necrosis. Oxidation at Met-46 may play a pivotal role in the formation of these insoluble structures. This modification has been detected in vitro following treatment with free radical donor (+/-)-(E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide. It has been proposed to destabilize nearby residues, increasing the likelihood of secondary oxidative damages, including oxidation of Tyr-45 and Met-105. This cascade of oxidations may augment GAPDH misfolding, leading to intermolecular disulfide cross-linking and aggregation. |
BioGrid: | 108868.233 interactions. |
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各位老师,在用凝集法或去溶剂法制备BSA的纳米微球,查阅相关文献,使用100mg的BSA溶于10ml的去离子水中,配成1wt%的水溶液,然后在磁力搅拌下,以1ml/min或2ml/min的速度加入乙醇40ml。在加入过程中,溶液变成乳白色,然后在加入就逐渐变清凉,但同时有沉淀出现,请问各位实验过程有啥问题啊?采取哪些措施可以避免出现沉定?另外制备纳米颗粒的过程,对BSA有具体的要求吗
我理解就是一个是包裹,一个是镶嵌
1.微囊的形态与粒径及分布
2.微囊的载药量与包封率
3.微囊药物的释放速率
4.有机溶剂残留量
微囊与微球的载体材料
常用的载体材料:
1.天然高分子材料
(1)明胶
明胶是由氨基酸与肽交联形成的直链聚合物。
明胶分酸法明胶(A型)和碱法明胶(B型)。A型明胶等电点为7~9,B型明胶稳定而不易长菌,等电点为4.7~5.0。两者的成囊性无明显差别,作囊材的用量为20~100g/L 。
可生物降解,几乎无抗原性。
(2) 阿拉伯胶
一般常与明胶等量配合使用,作囊材的用量为20~100g/L,亦可与白蛋白配合作复合材料。
(3) 海藻酸盐
系多糖类化合物,常用稀碱从褐藻中提取而得。海藻酸钠可溶于不同温度的水中,不溶于乙醇、乙醚及其它有机溶剂;不同Mav产品的粘度有差异。可与甲壳素或聚赖氨酸合用作复合材料。因海藻酸钙不溶于水,故海藻酸钠可用CaCl2固化成囊。
(4) 壳聚糖
壳聚糖是由甲壳素脱乙酰化后制得的一种天然聚阳离子型多糖,可溶于酸或酸性水溶液,无毒、无抗原性,在体内能被溶菌酶等酶解,具有优良的生物降解性和成膜性,在体内可溶胀成水凝胶。
2.半合成高分子材料
作囊材的半合成高分子材料多系纤维素衍生物,其特点是毒性小、粘度大、成盐后溶解度增大。
(1) 羧甲基纤维素盐(CMC-Na)
常与明胶配合作复合囊材,一般分别配1~5g/L CMC-Na及30g/L明胶,再按体积比2:1混合。CMC-Na遇水溶胀,体积可增大10倍,在酸性液中不溶。水溶液粘度大,有抗盐能力和一定的热稳定性,不会发酵,也可以制成铝盐CMC-A1单独作囊材。
(2)醋酸纤维素酞酸酯(CAP)
在强酸中不溶解,可溶于pH>6的水溶液,在二氧六环、丙酮中溶解,水、乙醇中不溶。用作囊材时可单独使用,用量一般在30g/L左右,也可与明胶配合使用。
(3)乙基纤维素(EC)
化学稳定性高,适用于多种药物的微囊化,不溶于水、甘油或丙二醇,可溶于乙醇,易溶于乙醚,遇强酸易水解,故对强酸性药物不适宜。用乙基纤维素为囊材时,可加入增塑剂改善其可塑性。
(4)甲基纤维素(MC)
在水中溶胀成澄清或微浑浊的胶体溶液,在无水乙醇、氯仿或乙醚中不溶。用作囊材的用量为10~30g/L,亦可与明胶、CMC-Na、聚维酮(PVP)等配合作复合囊材。
(5)羟丙甲纤维素(HPMC)
冷水中能溶胀成澄清或微浑浊的胶体溶液,pH值4.0~8.0(1%溶液,25℃) ,无水乙醇、乙醚 或丙酮中几乎不溶。
3.合成高分子材料
有生物不降解的和生物降解的两类。
生物不降解、且不受pH影响的囊材有聚酰胺、硅橡胶等。
生物不降解、但可在一定pH条件下溶解的囊材有聚丙烯酸树脂类、聚乙烯醇等。
生物降解的材料:聚碳酸酯、聚氨基酸、聚乳酸(PLA)、乙交酯丙交酯共聚物(PLGA)、聚乳酸-聚乙二醇嵌段共聚物(PLA-PEG)ε-己内酯与丙交酯共聚物等。特点:无毒、成膜性好、化学稳定性高,可用于注射。
聚酯类是迄今研究最多、应用最广的生物降解的合成高分子,它们基本上都是羟基酸或其内酯的聚合物。
常用的羟基酸是乳酸(1actic acid)和羟基乙酸(glycolic acid)。乳酸缩合得到的聚酯称聚乳酸,用PLA表示,由羟基乙酸缩合得的聚酯称聚羟基乙酸,用PGA表示;由乳酸与羟基乙酸缩合而成的,用PLGA表示,亦可用PLG表示。有的共聚物经美国FDA批准,也作注射用微球、微囊以及组织埋植剂的载体材料。
我最近用乳化交联法制备壳聚糖盐酸盐微球,药物是水不溶性(醋酸共晶体),将药物乙醇溶液加入到3%壳聚糖盐酸盐溶液中,混合均匀,加入到含2%span80的液体石蜡,油水比6:1,乳化半小时,加入1.5ml戊二醛,交联30min,离心,用石油醚,丙酮各洗两次,干燥,得微球。
但是我研磨微球,用乙醇溶解药物,超声3h,在紫外下根本检测不到药物,包封率就没法算,我测了药物在石油醚和丙酮中都有一定的溶解度,洗的时候溶剂层也有药物的颜色,是不是微球中的药物都被洗了出来?还是只是把游离的药物洗了出来?药物根本就没被包进去?但是药物在有机溶剂中都有一定的溶解度,我该怎么做?有大神愿意告知一二吗????
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