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| application information |
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| Recommended dilution | 1:2000 with ECL (WB) |
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| Expected | apparent MW | 27.8 | 27.5 kDa |
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| Confirmed reactivity | Arabidopsis thaliana, Hordeum vulgare, Nicotiana tabacum, Solanum lycopersicum (with higher load /well), Spinacia oleracea | |
| Predicted reactivity | Arthrospira platensis, Beta vulgaris, Glycine soja, Gossypium arboreum, Medicago truncatul, Morus notabilis, Oryza sativa, Phaseolus vulgaris, Riciunus communis, Theobroma cacao, Zea mays, Zostera marin, for reactivity on algae - please inquire | |
| Not reactive in | no confirmed exceptions from predicted reactivity are currently known |
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| Additional information | ||
| Selected references | to be added when available, antibody released in December 2015 | |
application example
5 µg of total protein from Spinacia oleracea (1), Arabidopsis thaliana (2), Solanum lycopersicum (3), Nicotiana tabacum (4), Hordeum vulgare (5) leafs were separated on 12 % SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with 5 % milk in TBS for 1h at room temperature (RT) with agitation. Blot was incubated in the primary anti-S5 antibody at a dilution of 1: 2 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 15 seconds.
RPS5 antibodies will react with Solanum lycopersicum when load per well is increased to 10-15 µg/well.
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制备抗体都需要进行纯化处理
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.
清冷冻干燥后保存。
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。
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
