Recombinant Human Pro-SUMO3 Protein, CF Summary
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins.Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration.The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard.In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
UL-760
Shipping | The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below. |
Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
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Reconstitution Calculator
Background: SUMO3
Human Small Ubiquitin-like Modifier 3 (SUMO3), also known as SMT3A, is synthesized as a 103 amino acid (aa), propeptide with a predicted 11.5 kDa. SUMO3 contains a two aa C-terminal prosegment. Human SUMO3 shares 83% sequence identity with mouse SUMO3. SUMO3 also has high aa sequence homology to SUMO2 and SUMO4, 87% and 75%, respectively. SUMO3 shares only 47% sequence identity with SUMO1. SUMOs are a family of small, related proteins that can be enzymatically attached to a target protein by a post-translational modification process termed SUMOylation (1-3). All SUMO proteins share a conserved Ubiquitin domain and a C-terminal diglycine cleavage/attachment site. Following prosegment cleavage, the C-terminal glycine residue of SUMO3 is enzymatically attached to a lysine residue on a target protein. In humans, SUMO3 is conjugated to a variety of molecules in the presence of the SAE1/UBA2 SUMO-activating (E1) enzyme and the UBE2I/Ubc9 SUMO-conjugating (E2) enzyme (4,5). In yeast, the SUMO-activating (E1) enzyme is Aos1/Uba2p (6). Because of the high level of sequence homology most studies report effects of SUMO2/3. For example, addition of SUMO2/3 was shown to modulate the function of ARHGAP21, a RhoGAP protein known to be involved in cell migration (7). Other reports indicate that the conjugation by SUMO2/3, but not SUMO1, may represent an important mechanism to protect neurons during episodes of cerebral ischemia (8,9). However, studies suggest that SUMO2/3 expression is regulated in an isoform-specific manner since oxidative stress downregulated the transcription of SUMO3 but not SUMO2 (10).
All SUMO isoforms are translated with additional C-terminal residues that have to be removed to generate the active protein. Pro-SUMO-3 (103 aa) is the inactive precursor of SUMO-3 (92 aa) and is processed at the C-terminus by SUMO-3 specific proteases (SENPs).The resulting SUMO-3 protein has the conserved C-terminal Gly-Gly residues that function in activation and conjugation reactions. This protein can be used as a negative control in SUMOylation reactions or as a substrate for SENPs. NCBI Accession # NM_006936.
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protocol如下:
灌注取脑后,4%多聚甲醛4℃固定24-48h。修块3-4mm。
脱水(常温):45%、55%、65%、75%、85%、95%、100%酒精各30min;
透明(常温):(100%酒精+二甲苯)/210min,二甲苯①20min,二甲苯②至完全透明(约20min)
浸蜡(60℃):(二甲苯+石蜡①)/210min,石蜡①30min,石蜡②40min;石蜡①为低熔点,石蜡②为高熔点
包埋:浸蜡后用石蜡②手动包埋;
切片厚度:6um、(8um也切的有)
展片:40℃水中展片;
烤片:60摄氏度烤片机侧烤约5min后吸去水分,转移至60℃温箱烤约4h;
脱蜡:烤片后趁蜡未凝固,进行脱蜡。二甲苯5min×2,100%酒精3min,95%酒精2min,
85%酒精2min,75%酒精2min,流水冲洗2min(放在缸子里用水泡的,换了几次水);
染色:
HE染色:苏木精5-8min(新配为5min,时间长可为8min,用前过滤),流水冲洗3min
1%盐酸酒精分化5s,流水冲洗1min,0.1%伊红1-2min,
脱水:85%酒精2min,95%酒精2min,100%酒精2min,
100%无水乙醇:二甲苯(1:1)2min
透明:二甲苯(I)3min,二甲苯(II)3min;
CV染色:浸入CV染液中10-20min,
脱水:75%乙醇2min,85%乙醇2min,95%乙醇2min,100%无水乙醇2min,
100%无水乙醇:二甲苯(1:1)2min
透明:二甲苯(I)3min,二甲苯(II)3min;
封片:中性树胶封片(整个染色过程避免干片)。
部分HE染色、CV染色片子结果如图所示,基本上没有完整的片子(染色时,片子随着在液体里的时间脱片、烂片的程度加深),另CV染色着色浅。
请教战友是什么原因?哪里出了问题,该怎么解决?
尤其是前面两个,后面的还可以从字面理解着做,前面这两个就纯把我搞晕菜了.
全量法:测水浸出物的
酒石酸铁比色法;
紫外分光光度法;
恒重法:测水含量的
谢谢谢谢
[国家标准物质网]高频红外碳硫分析仪是一种常用的分析仪器,能快速、准确地测定钢、铁、合金、有色金属、水泥、矿石、玻璃及其它材料中碳、硫两元素的质量分数。高频红外碳硫分析仪在使用过程中需要进行定期维护,今天我们就来具体介绍一下高频红外碳硫分析仪的日常维护方法。
高频红外碳硫分析仪日常维护要点
1、高频红外碳硫分析仪分析样品所用的坩埚需要马弗炉中加热温度升到1000度到1200度,恒温2小时后,自然冷却至一定温度时,取出放干燥器中冷至室温备用。
2、分析低含量样品所用的钨粒需在恒温箱中,加热至200度烘2小时后,取出在干燥器中冷却后使用。
3、高频红外碳硫分析仪设置了高频输出功率调节,在未打开仪器电源之前,用万用表测试电网电压后,将换档开关拨至电网电压基本一致的档位上,开始调试仪器。方法是当然界电网电压为220V时,调至230V档为减小输出功率,调至210档为增加输出功率。
4、打开总氧气阀门,打开仪器电源,按技术员所教操作。
5、每天仪器示打开电源前或分析200个样品后,需清扫一次过滤网及整个炉腔。
6、每天应检查高频炉正前方的干燥剂和脱脂棉。一旦发现干燥剂1/3变红或结块,都应立即更换。脱脂棉长期使用1/3变黄后,也应立即更换。更换时将旋转试管下方的螺杆,使试管下降至能离开上端面为止,取出试管,更换后按原装好拧紧,检查不漏气为止。
7、每月应清洗过滤网一次,清洗方法是:按上述“清扫方法”取出过滤网后,再取出过滤网上、下两只密封圈,将粉尘清扫后,放入超声波清洗器中,在清洗槽内加入蒸馏水和洗洁精。打开超声波清洗器电源。30分钟后取出过滤网,用蒸馏水反复清洗后,用无水乙醇清洗。最后取出过滤网用电吹风干或放入烘箱烘干,装上、下O型密封圈,涂上真空硅脂,按原位老妈子,检查不漏气为止。
8、高频红外碳硫分析仪使用时每季度应检查经过压紧阀内部的硅橡胶管,若发现有老化或弹性不好时应立即更换。
摘自:国家标准物质网
谁用过亲水性改性的聚丙烯膜养过细胞,最好是whatman的,效果怎么样?十分感激!:)
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新帖要有题目,且要尽量展示帖子主要内容意思~bywzmc225
本版发帖须知及新手指南
主药:50mg
mcc:20mg
l-hpc:10mg
cms-na:10mg
5%pvp50%乙醇qs
外加
ms:0.5mg
cms-na:3mg
50度干燥
以上处方不粘冲,但因上处方的辅料与主药有相互作用,导致片子变黄
现换成以下处方
处方1
主药:50mg
预胶化淀粉:20mg
淀粉:20mg
8%淀粉浆:qs
外加
ms:0.5或1mg
滑石粉:3或5mg
60度干燥
以上处方粘冲,为何?
处方2
主药:50mg
预胶化淀粉:20mg
淀粉:10mg
甘露醇:10mg
8%淀粉浆:qs
外加
ms:0.5或1mg
滑石粉:3或5mg
60度干燥
以上处方粘冲,为何