专家观点
哥本哈根大学Dr.John Couchman认为优质抗体,尤其是多抗,应该是2步纯化:
Not only must I affinity purify the antibody but also perform antigen
adsorption on my tissues sections. Unfortunately, many commercial antibodies
are not affinity purified from serum.
哈佛大学神经学院的主任Dr. Clifford Saper在公开发表的文章中指出经过抗原亲和纯化
的抗体的特异性更好:
Affinity purify antisera by running them over a column to which the antigen is
fixed to a support matrix. Such antisera are less prone to artifact and tend
to have lower background staining.
专家观点
科研人员认为对于那些不提供多肽信息的抗体,科学界是应该抵制的。
The antigen is not clearly stated。 Sometimes for “proprietary” reasons,
details on the antigen are withheld. This is not good science, and we should
boycott those antibodies.
依然是哈佛大学研究人员的观点:使用那些没有多肽信息的抗体是难以得到可重复的实验
结果,也不适合于严谨的科学研究。
Several manufactures have claimed that sequence information on the antigen was
“proprietary,” and they would not provide it. Because work using these
antibodies is inherently not repeatable, such papers are not acceptable for
publication.
Some companies are well-known for keeping the exact sequence of the immunizing
peptide as a “trade secret”. Investigators should be aware that such
antisera are not fit for use in serious scientific work.
哈佛大学的Clifford Saper教授在公开发表的文章中更是直接指出:
永远不要购买没有抗原信息的抗体,这样的研究成果既不科学,也不能被发表!
Never buy antibodies for which the manufacturer will not disclose the
structure of the immunizing antigen. These reagents are not fit for scientific
work, and the work you do with them will not be publishable.
专家观点
抗体所识别的蛋白,可能并不是免疫动物是所使用的的抗原蛋白。
Antibodies are biological agents, not standard chemical reagents. Antibodies
may bind to a wide variety of antigens other than the one that they were
raised to recognize.
专家观点
兔子因为体积小,每次产生抗体的量是极其有限的,一方面对于同一只兔子,因为不同的
免疫时间,所以产生的抗体是有差异的;另一方面即使在同一个免疫时间,不同兔子因为
个体的差异,产生的抗体也是不一样的。因此,相比较兔子,山羊来源的抗体显然质量更
稳定,批次之间的差异更小。
Because bleeds from rabbits are limited in volume, a given manufacturer may
run out of a particular bleed, and substitute another bleed from the same
animal, or a bleed from a difference animal …… Quite often the new bleeds
have not been checked to make sure that they stain the same thing.