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| application information |
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| Recommended dilution | 1: 8000 (ELISA), 1: 1000 with standard ECL (WB) |
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| Expected | apparent MW | 25.9 kDa |
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| Confirmed reactivity | Arabidopsis thaliana |
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| Predicted reactivity | ||
| Not reactive in | no confirmed exceptions from predicted reactivity known in the moment |
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| Additional information | Protein or membrane sample should be treated at 70°C for 10 min before loading on the gel. Diluted antibody solution can be used 2 to 3 times within one month if it contains 0.1 % sodium azide as preservative and is stored at -20ºC to -80ºC. Manufactured by Operon Biotechnologies. |
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| Selected references | Ishikawa et al. (2005). Novel type aquaporin SIPs are mainly localized to the ER membrane and show cell-specific expression in Arabidopsis thaliana. FEBS Lett. 579:5814-5820. | |
application example
crude membrane fraction/lane from Arabidopsis thaliana stems (1,3) and crude membrane fraction from yeast cells expressing SIP2;1 (2,4) were separated on 12 % SDS-PAGE and blotted 1h to PVDF membrane (40 min. at 10 V using BioRad semidry transfer). Filters were blocked 1h with 5 % low-fat milk powder in TBS-T (0.05% Triton X.100). Membranes were washed 5 times with TBS-T, each time in a fresh polystyrene box and probed with anti-AtSIP2;1 antibodies (AS09 496, 1:1000, 1h) and secondary anti-rabbit (1:2000, 1 h). All steps were performed in RT with agitation. Peptide used to elicit anti-SIP2;1 antibodies have been incubated with antibodies - right panel. |
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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
