| Clone | Polyclonal |
| Units | 1 ml |
| Host | Goat |
| Application | immunoelectrophoresis |
BackgroundIn immunoelectrophoresis against rabbit milk precipitation of not less than 8 different proteins may be obtained. However the number of visible precipitin lines varies widely among individual specimens of rabbit milk. The total concentration of protein, their proportion of the different protein components change markedly during the course of lactation. A protein detected in a rabbit secretion by this antiserum may be of secretory origin, or it may be a plasma protein presumably entering in the secretion from the surrounding capillary vessels and through interstitial tissues. Virtually every plasma protein may also be present at a detectable level in milk. Not all precipitable proteins have been identified. Known proteins include secretory IgA and other immunoglobulins, free secretory component, lactoferrin, components of the complement system and albumin. In immunoelectrophoresis to identify the protein pattern in an individual milk sample, to compare individual patterns and to make comparisons with serum protein patterns or those of other secretions; to identify individual components, and significant changes in concentration. To test the purity of an isolated milk protein.
SourcePooled whole rhesus rabbit milk. Freund’s complete adjuvant is used in the first step of the immunization procedure.
ProductDelipidated, heat inactivated, lyophilized, stable whole antiserum No preservative added. Total protein and IgG concentrations in the antiserum are comparable to those of pooled normal goat serum. No foreign proteins added. Reconstitute the lyophilized antiserum by adding 1 ml sterile distilled water.
Formulation: Delipidated, heat inactivated, lyophilized, stable whole antiserum No preservative added. Total protein and IgG concentrations in the antiserum are comparable to those of pooled normal goat serum. No foreign proteins added.
SpecificityPrecipitating polyclonal Goat antiSerum to Rabbit milk proteins.
Species Reactivity: Inter-species cross reactivity is a normal feature of antibodies to animal proteins since homo- logous proteins of different species frequently share antigenic determinants. This antiSerum has not been adsorbed for such cross reactivity. Consequently it is not species-specific.
ApplicationsPrecipitation assays. In immunoelectrophoresis use 2 μl serum or equivalent against 120 μl antiserum. In double radial immunodiffusion (Ouchterlony) use a rosette arrangement with 10 μl antiserum in 3 mm diameter center well and 2 μl serum samples (neat and serially diluted in 2 mm diameter peripheral wells. Different bleedings of the immunized animals are pooled to obtain a broad spectrum balanced against the varying concentrations of the individual serum protein components.
StorageThe lyophilized antiserum is shipped at ambient temperature and may be stored at +4°C; prolonged storage at or below -20°C.Dilutions may be prepared by adding phosphate buffered saline (PBS, pH 7.2). Repeated thawing and freezing should be avoided. If a slight precipitation occurs upon storage, this should be removed by centrifugation. It will not affect the performance of the antiserum.Diluted antiserum should be stored at +4°C, not refrozen, and preferably used the same day.
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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2.加大辣根过氧化物酶标记的链霉亲和素；
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！！
请大家帮帮忙，第一步加完抗原后必须封闭吗
目前，我用该蛋白的鼠源性单抗来包被ELISA板子，5%BSA37度封闭二小时后，洗涤四次后将板子分三个组加入抗原：其中一个组中加入的是我重组的蛋白，另一个组为用其它方法证实含该蛋白的人体内血清，最后一个组加入PBS。37度一小时后洗涤四次后每孔加入我制备的兔源性多抗。37度一小时洗涤六次后我再加入酶标记的抗兔的抗体37度40分钟后洗涤六次后TMB显色。
问题就在我显色后所有的孔都是阳性，其中以抗原为我的重组蛋白组与PBS组OD值最高，两组都可以达到1.8以上，而加入的抗原为人体血清组低，为0.6-1.0不等。
后面我优化条件，封闭液用过5%的脱脂奶粉，2%的BSA，包被的单抗浓度摸了梯度，加入的多抗也摸了梯度，酶标的抗体也摸了梯度。结果都没有改变，连趋势每次都是一样以重组蛋白组与PBS组OD最高。
今天我还试着把抗原换成其它蛋白也做出了强阳性？
请教各位高手，我的ELISA该如何往下面做了啊，是不是我的抗体本身有问题呢？
对了，我包被用的单抗是用GE公司的预装柱纯化的，很纯，在考染中是看不到一条杂带的。
ELISA双抗体夹心法(enzyme linked immunosorbent assay——sandwich technique)的原理是将特异性抗体结合到固相载体上形成固相抗体，然后和待检血清中的相应抗原结合形成免疫复合物，洗涤后再加酶标记抗体，与免疫复合物中抗原结合形成酶标抗体-抗原-固相抗体复合物，加底物显色，判断抗原含量。
生物帮有相关介绍。编码RNA http://doc.bio1000.com/show-3399.html
（2）封闭：将已包被的板用洗涤液洗涤2次，每孔加入120 μL封闭液，封闭一段时间，取出，甩干备用。
（3）竞争反应：在制备好的板中每孔加入50 μL系列浓度的标准品溶液和50 μL稀释好的酶标抗体溶液，孵育一段时间。
（4）洗板：用洗涤液洗涤5次。
（5）显色反应：每孔加入显色溶液100 μL，孵育一段时间。
（6）终止：每孔各加入终止液，在酶标仪上测定各孔的吸光值。
