该单克隆抗体用与人类BiP中Gly584位点附近的氨基酸序列、人ERp44序列、人ERp72中Met279位点附近的氨基酸序列对应的人工合成肽段免疫动物而制成。该多克隆抗体用与人ERp57中Gly117位点附近、人Grp94中Met622位点附近和人Ero1-Lα中Leu218位点附近序列对应的人工合成肽段免疫动物而制成。该抗体使用蛋白A和蛋白亲和层析纯化而得。ER Protein Folding Antibody Sampler Kit包含了研究细胞内翻译起始的试剂。该试剂盒包含足够每个一抗完成四次Western blots的一抗和二抗。分泌蛋白合成后转位到内质网进行翻译后修饰和正确折叠。为了形成其天然构象,很多分泌蛋白需要形成分子间或分子内二硫键(1)。这个过程成为氧化蛋白折叠。二硫键异构酶(PDI)催化蛋白质天然活性形成的二硫键形成和异构变化(2)。对氧化折叠的研究提示分子氧可以氧化内质网蛋白质Ero1,Ero1反过来通过二硫键交换氧化PDI(3)。之后PDI催化的二硫键在正在折叠的蛋白质上形成(3)。其它驻内质网的拥有硫氧还蛋白同源性区域的蛋白,包括内质网应激蛋白72、57和44(ERp72, ERp57和ERp44),组成了PDI家族(4,5,6)。内质网也包含一组分子伴侣,包括Grp94,以帮助蛋白质正确折叠。Grp94是葡萄糖调节蛋白(7),与Hsp90有序列同源性(8)。BiP是另一种分子伴侣,当蛋白质正确折叠被干扰时表达增加。BiP结合到错误的蛋白质上,防止其形成聚集物,并帮助其重新正确折叠(9)。
原厂资料:
Specificity / Sensitivity
Each antibody in the ER Protein Folding Antibody Sampler Kit detects endogenous levels of its target protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Gly584 of human BiP, the sequence of human ERp44, the residues surrounding Met279 of human ERp72 protein and the sequence of human PD1. Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequences around Gly117 of human ERp57, Met622 of human Grp94 and Leu218 of human Ero1-Lα. Polyclonal antibodies are purified by protein A and peptide affinity chromatography.
Description
The ER Protein Folding Antibody Sampler Kit contains reagents to investigate the initiation of translation within the cell. The kit contains enough primary and secondary antibodies to perform four Western blot experiments per primary antibody.
Background
After their synthesis, secretory proteins translocate into the endoplasmic reticulum (ER) where they are post-translationally modified and properly folded. To reach their native conformation, many secretory proteins require the formation of intra- or inter-molecular disulfide bonds (1). This process is called oxidative protein folding. Disulfide isomerase (PDI) catalyzes the formation and isomerization of these disulfide bonds (2). Studies on mechanisms of oxidative folding suggest that molecular oxygen oxidizes the ER-protein Ero1, which in turn oxidizes PDI through disulfide exchange (3). This event is then followed by PDI-catalyzed disulfide bond formation on folding proteins (3). Other ER resident proteins that possess the thioredoxin homology domains, including endoplasmic reticulum stress proteins 72, 57 and 44 (ERp72, ERp57 and ERp44), constitute the PDI family (4,5,6). The ER also contains a pool of molecular chaperones, including Grp94, to help proteins fold properly. Grp94 is a glucose-regulated protein (7) with sequence homology to Hsp90 (8). BiP is another chaperone whose synthesis is increased when protein folding is disturbed. BiP binds to misfolded proteins to prevent them from forming aggregates and assists in proper refolding (9).
注意事项:
Storage: Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM
NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not
aliquot the antibodies.