
Units | 0.1 ml |
Host | Rabbit |
Application | ELISAIFAIHC(P)ImmunofluoresenceImmunohistochemistry (paraffin)RadioimmunoassayRIAWestern Blotting |
BackgroundType I Collagen usually exists as a heterotrimer formed by alpha 1(I) and alpha 2(I) chains and is found in bone, cornea, skin and tendon. In foetal tissues also homotrimers of alpha-1(I) are found, but they are not constituents of normal adult tissues. Collagens consist of a family of highly specialized glycoproteins of which at least 16 genetically distinct types are known to date. The basal unit of a collagen molecule consists of a triple-helical structure formed by 3 alpha-chains. Predominant amino acids are glycine, proline and hydroxproline. Regularly also lysines and hydroxylysines occur, which are responsible for cross-linkage and glycosylation of the protein chains. Different composition of alpha-chains and different glycosylation contribute to the high variability of collagens in different tissues and organs. Tunafish collagen type I.
Source
Immunogen: Purified collagen type I from tuna fish skin
Productaffinity purified antibody lyophilized from phosphate buffered solution; no BSA and preservative added!
Purification Method: affinity purified antibody lyophilized from phosphate buffered solution; no BSA and preservative added!
Concentration: app. 1 mg/ml
Secondary Reagents: Anti-rabbit IgG-conjugates, e.g. anti-rabbit IgG:FITC (Art. No. FI-1000) or anti-rabbit IgG:DyLight488 (Art. No. DI-1488).
Specificity
Species Reactivity: Tunafish, crossreactivity with collagen type I of other species <0.1%>0.1%>
ApplicationsIHC(P), IFA, ELISA, RIA, IB/WB
Incubation Time: IHC(P) 60 min at RT or 2-8°C over night
Working Concentration: (purified, lyophilized) IFA ? 1:80, IHC(P) ? 1:500, ELISA ? 1:200 (OD ? 500)
Pre-Treatment: After de-waxing the tissue slices they are treated with 0.2% hyaluronidase (app. 300 U/mg e.g. Art. No. HYA02-50) in TBS 15 min at 37°C. Thereafter non-specific binding is blocked by blocking serum or 3% BSA in TBS. For peroxidase systems blocking with 1% peroxide solution in TBS for 30 min at RT is recommended.
Positive Control: Tunafish skin
Storage-20°C
CautionThis product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. It may contain hazardous ingredients. Please refer to the Safety Data Sheets (SDS) for additional information and proper handling procedures. Dispose product remainders according to local regulations.This datasheet is as accurate as reasonably achievable, but Exalpha Biologicals accepts no liability for any inaccuracies or omissions in this information.
References1. Rigo C., Hartmann D.J., Bairati A. - Electrophoretic and immunochemeical study of collagens from Sepia officinalis cartilage. Biochim. Biophys. Acta 2002, 1572, 77-84.
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一般来说多克隆的阳性率高一些,但出现假阳性的比例也高一些。
制备抗体都需要进行纯化处理
抗体的特异性鉴定 抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在 IC50时的浓度,并按下列公式计算交叉反应率。 如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示 这一抗血清与其他抗原物质的交叉反应率近似为 0,即该血清的特异性较好。
抗体的亲和力 是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小、抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键、疏水键、侧链相反电荷基因的库仑力、范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol,通常K的范围在 108 ~1010 /mol,也有多达 1014 /mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。向左转|向右转
又由于自然存在的抗原大都存在多个抗原表位,会刺激机体产生多种针对同一抗原的不同抗原表位相应的不同抗体.

