- Overview
- Data/Specifications
- Literature/Support
- How To Use
- Related Products
Overview
Type I collagen is the most abundant collagen and is found in connective tissues including tendon, ligament, dermis and blood vessel. It is the major component and the primary determinant of tensile strength of the extracelluar matrix (ECM). It is widely used as a thin layer on tissue-culture surfaces to enhance the attachment and proliferation of a variety of cells including endothelial cells, fibroblasts, hepatocytes, epithelial cells etc. In addition, collagen I can self-assemble into a 3-D superamolecular gel in vitro, making it an ideal biological scaffold to promote more in vivo-like cellular morphology and function.
Type III collagen is the second most abundant collagen in tissues and is found most commonly in tissues exhibiting elastic properties such as skin, lungs, intestinal walls and walls of blood vessels. It is a homotrimer comprised of three alpha-1 chains and resembles other fibrillar collagens in structure and function. It is synthesized as procollagen, similary to collagen I, but the N-terminal propeptide remains attached in the mature fibrillar type III form.
Symbols/Related Terms:
- COL1A1
- COL1A2
- COL3A1
- Collagen type III, alpha 1
- osteogenesis imperfecta
Data/Specifications
Purity:
Bovine collagen type I - 45%
Bovine Collagen type III - 45%
Bovine collagen other types - below 10%
Non-collagen proteins - below 0.5%
Form: 10 mg lyophilized, salt-free
Purification: Partial pepsin digestion in acidic conditions and differential salt precipitation.
Source: Bovine (calf) skin. Collagen was extracted from washed dissected tissue into dilute acetic acid after pepsin treatment. Collagen type I and III were purified by using differential salt precipitation.
Reconstitution: Use 0.5 M acetic acid, pH 2.5. Dissolved collagen retains immunologic properties of native collagen. Structure of native collagen confirmed by ability to form microfibrils.
Storage:Collagen dissolved in acetic acid is stable at 4 °C for 1 month. Lyophilized collagen long term storage (2 years) at -20°C or lower.
Literature/Support
Bovine Type I and III Collagen Insert (PDF)
Note: inserts are for review only. Please refer to the insert shipped with your product to ensure the most up-to-date revision is being used.
How To Use
Key Applicatons:
- Coating material for cell culture studies.
Note: May not be suitable for 3-D gel formation.
Related Products
ebiomall.com
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选用和特异性抗体同种亚型的，同样荧光素标记的抗体，采用同样的染色步骤染同样浓度的细胞。然后上机检测。如果细胞有非特异性的结合，那同型对照的荧光强度可能会比未染色的阴性对照要高一些。
同型对照只用于那种阳性和阴性之间没有明确分解的情况。如果阳性细胞群非常清楚，是不需要同型对照的。另外，在多色同染的情况下，也不采用同型对照，而是采用FMO（fluorensence minus one）对照。
另外，对于同型对照的使用，也有很多反对意见。就算是同型抗体，也可能会有不同的亲和力，并不一定就能真实反应非特异性的结合。
作用：(1)特异性结合抗原：抗体本身不能直接溶解或杀伤带有特异抗原的靶细胞,通常需要补体或吞噬细胞等共同发挥效应以清除病原微生物或导致病理损伤.然而,抗体可通过与病毒或毒素的特异性结合,直接发挥中和病毒的作用. (2)激活补体：IgM、IgG1、IgG2和IgG3可通过经典途径激活补体,凝聚的IgA、IgG4和IgE可通过替代途径激活补体. (3)结合细胞：不同类别的免疫球蛋白,可结合不同种的细胞,产生不同的疚,参与免疫应答. (4)可通过胎盘及粘膜：免疫球蛋白G(IgG)能通过胎盘进入胎儿血流中,使胎儿形成自然被动免疫.免疫球蛋白A(IgA)可通过消化道及呼吸道粘膜,是粘膜局部抗感染免疫的主要因素. (5)具有抗原性：抗体分子是一种蛋白质,也具有刺激机体产生免疫应答的性能.不同的免疫球蛋白分子,各具有不同的抗原性. (6)抗体对理化因子的抵抗力与一般球蛋白相同：不耐热,60～70℃即被破坏.各种酶及能使蛋白质凝固变性的物质,均能破坏抗体的作用.抗体可被中性盐类沉淀.在生产上常可用硫酸铵或硫酸钠从免疫血清中沉淀出含有抗体的球蛋白,再经透析法将其纯化. (7)通过与细胞Fc受体结合发挥多种生物效应 ①调理作用 IgG、IgM的Fc段与吞噬细胞表面的FcγR、FcμR结合,增强其吞噬能力,通常将抗体促进吞噬细胞吞噬功能的作用称为抗体的调理作用 (opsonization). ②发挥抗体依赖的细胞介导的细胞毒作用
2、对人体疾病而言，按照经典且目前行之有效的疫苗和免疫学角度看，中和抗体对预防这类疾病更有效。但很多资料也表明，可能那些non-neutralizing但具有其他作用的功能性抗体在控制和预防感染性疾病中也发挥着十分重要的作用，例如补体作用，ADCC，ADCVI等等。
先说骨髓瘤细胞和B淋巴细胞融合，共有3种情况，B淋巴和B淋巴融合，B淋巴和骨髓瘤细胞融合，骨髓瘤细胞和骨髓瘤细胞融合
第一次筛选是为了筛选出能无限增殖的细胞，这只有两种情况：
1、经过免疫的B淋巴细胞和骨髓瘤细胞的融合细胞
2、骨髓瘤细胞和骨髓瘤细胞的融合细胞
第二次是为了筛选出能产生单克隆抗体的细胞
那么只可能是，经过免疫的B淋巴细胞和骨髓瘤细胞的融合细胞。
所以有两次筛选
