Clone | Polyclonal |
Units | 1 ml |
Host | Rabbit |
BackgroundThe reactivity of the antiserum is directed to the Fab subunits of intact IgG, IgA IgM and other Ig classes of both light chain types, with their Fab or F(ab’)2 subunits, and with free light chains of kappa and lambda type. In immunoelectrophoresis and double radial immunodiffusion using various antiserum concentrations against pooled monkey serum a characteristic precipitin line is obtained, which shows a reaction of identity with the precipitin line of the purified Fab. Direct staining of fixed cell and tissue substrates; to demonstrate the intracellular presence of free or Ig-bound subunits of both kappa or lambda type. In general this conjugate is not recommended as direct or indirect screening reagent for If isotypes on surface membranes of vital lymphoid cells. The activity to the common Ig/Fab subunit may result in the staining of immunoglobulins bound to the Fc-receptors on non-lymphoid cells. Combinations of isotype-specific reagents should be used instead for this purpose. This immunoconjugate is not pre-diluted. The optimum working dilution of each conjugate should be established by titration before being used. Excess labelled antibody must be avoided because it may cause high unspecific background staining and interfere with the specific signal.
SourcePurified normal Fab of polyclonal rhesus monkey IgG. Freund’s complete adjuvant is used in the first step of the immunization procedure.
ProductHorseradish peroxidase-coupled purified hyperimmune rabbit IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). No preservative added, as it may interfere with he antibody activity. It is reconstituted by adding 1 ml sterile distilled water, spun down to remove insoluble particles, divided into small aliquots, frozen and stored at or below -20°C.
Product Form: HRP
Formulation: Horseradish peroxidase-coupled purified hyperimmune rabbit IgG lyophilized from a solution in phosphate buffered saline (PBS, pH 7.2). No preservative added, as it may interfere with he antibody activity.
SpecificityHorseradish peroxidase–conjugated IgG fraction of polyclonal Rabbit antiSerum to Monkey Fab of IgG.
Species Reactivity: Inter-species cross-reactivity is a normal feature of antibodies to immunoglobulins, since immunoglobulins of different species frequently share antigenic determinants. Cross-reactivity of this immunoconjugate has not been tested in detail, however in double radial immunodiffusion (Ouchterlony) a reaction has been obtained with Human immunoglobulins.
ApplicationsELISA,Immunocytochemistry,Immunohistochemistry (paraffin),Dot blot,Immunoblotting.
StorageThe lyophilized conjugate is shipped at ambient temperature and may be stored at +4°C; prolonged storage at or below -20°C.Prior to use, an aliquot is thawed slowly at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the immunoconjugate.
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.
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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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