Clone | PEN-12 |
Isotype | IgG1 |
Units | 50 µg |
Host | Mouse |
Application | ELISAImmunohistochemistry |
BackgroundThis antibody is suitable for the detection of bound and free pentosidine in tissues and tissue extracts. Long-term incubation of proteins with glucose leads, through Schiff"s base and Amadori rearrangement products, to the formation of advanced glycation end products (AGE) which are characterized by fluorescence, brown color and inter- and intra-molecular cross-linking. Recent immunological studies using anti-AGE antibodies demonstrated the presence of AGE in (i) human lens, (ii) renal proximal tubules in patients with diabetic nephropathy and chronic renal failure, (iii) atherosclerotic lesions of arterial walls, (iv) ß2-microglobulin of carpal tunnel amyloid fibril deposits in patients with hemodialysis-related amyloidosis and (v) brain tissues of patients with Alzheimer’s disease. Pentosidine is one of the Maillard products, which were detected by Sell and Monnier (1989). Pentosidines have been shown to cross-link Arginine and Lysine residues. They were found in ß2-microglobulin of patients suffering from hemodialysis-related amyloidosis (Miyata et al., 1996). Reacts with pentosidine-HSA and free pentosidine in competitive ELISA.
Source
Immunogen: Pentosidine-HSA
ProductProtein G affinity purified antibody from ascites in stabilized buffer, containing 50% Block Ace? (Casein-containing solution, Dainippon Co.) and 0.1% ProClin? (Rohm & Haas) as a preservative
Purification Method: Protein G affinity purified antibody from ascites in stabilized buffer, containing 50% Block Ace? (Casein-containing solution, Dainippon Co.) and 0.1% ProClin? (Rohm & Haas) as a preservative
Concentration: 0.25 mg/ml
Secondary Reagents: We recommend the use of BIOLOGO"s Universal Staining System DAB (Art. No. DA005) or AEC (Art. No. AE005).
Specificity
Species Reactivity: not relevant
ApplicationsIHC, ELISA
Incubation Time: 60 min at RT or 18 hr at 2-8°C
Working Concentration: (liquid conc.) ELISA 0.1-0.5 µg/ml; IHC 2 µg/ml
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. Horiuchi S., Araki N., and Morino Y. (1991) Immunological approach to characterize advanced glycation end products of the Maillard reaction: Evidence for the presence of a common stucture. J. Biol. Chem. 266; 7329-7332. 2. Sell D.R. and Monnier V.M.(1989) : Structure Elucidation of a Senescence Cross-link from Human Extracellular Matrix. Implication of pentoses in aging process. J.Biol.Chem.264 : 21597-21602 3. Miyata T, Taneda S., Kawai R., Ueda Y., Horiuchi S., Hara M., Maeda K., and Monnier V.M. (1996) : Identification of pentosidine as a native structure for advanced glycation end products in ?2-microglobulin-containing amyloid fibrils in patients with dialysis-related amyloidosis. Proc.Natl.Acad.Sci.USA 93 : 2353-2358
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1.标准曲线的标准品是否一定要梯度稀释,为什么?我试过非梯度稀释的,也可以达到线性R2=0.99.
2.我用了CurveExpert做标曲,自动搜索后发现有10种提供的方程,各种形式的,其中一个十分适合我的实验结果(LogisticModel),而其他的感觉又不适合,因为结果常常为负值。这又是为啥捏?
3.实验的酶标仪最大OD值可以测到4,如果我的测量结果在1.3,是否像其他人所说的>1了就不准确了。
4.利用夹心法进行定量分析是否一定要使用线性方程?
不好意思啊,一下问了这么多问题,最近做了一个月的ELISA,完全摸不清头脑啊。谢谢各位了
我现在用商品化的试剂盒进行夹心法ELISA测定某抗体的浓度,盒子给定了4个标准品,标准品抗体浓度为70000~1000单位,测定步骤中要求对样品进行1:1000稀释后测定。
但是有的样本按照1:1000稀释后,最终OD值大于70000浓度的标准品的OD值,然后使用ELISACALC软件进行四参数拟合,超过了标准曲线的范围,就算不出来这个样品的浓度,但是如果在1000-70000之间的OD值就可以算出相应浓度。请教大神们,接下来我是否可以对样品进行1:2000,1:4000浓度稀释,算出结果后再×2、×4,算作此样品的浓度呢?还是直接给一个>70000的定性结果?或者可以有能算出来的其他软件?
Ps试剂盒中提及Dilutionlinearity(稀释线性?)为141%,这个是什么意思呢?
多谢!!
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请大家帮帮忙,第一步加完抗原后必须封闭吗
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