- SynonymComplement C5,C5,CPAMD4
- SourceHuman Complement C5, His Tag (CO5-H52Ha) is expressed from human 293 cells (HEK293). It contains AA Gln 19 - Cys 1676 (Accession # P01031-1).Predicted N-terminus: Gln 19 (β chain) & Thr 678 (α chain)Request for sequence
- Molecular Characterization
This protein carries a polyhistidine tag at the C-terminus.The mature form of Complement C5 is a disulfide-linked heterodimer composed of proteolytically cleaved α and β chain. Each α and β chain has a calculated MW of 73.9 kDa (β chain) and 114.4 kDa (α chain). The protein migrates as 66 kDa (β chain) and 110 kDa (α chain) under reducing (R) condition (SDS-PAGE) due to glycosylation.
- EndotoxinLess than 1.0 EU per μg by the LAL method.
- Purity
>90% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
Human Complement C5, His Tag on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
Immobilized Human Complement C5, His Tag (Cat. No. CO5-H52Ha) at 2 μg/mL (100 μL/well) can bind Anti-C5a (Human IgG1) with a linear range of 0.2-3 ng/mL (QC tested).
- BackgroundDerived from proteolytic degradation of complement C5, C5 anaphylatoxin is a mediator of local inflammatory process. C5 precursor is first processed by the removal of 4 basic residues, forming two chains, beta and alpha, linked by a disulfide bond. C5 convertase activates C5 by cleaving the alpha chain, releasing C5a anaphylatoxin and generating C5b (beta chain + alpha" chain). Activation of C5 by a C5 convertase initiates the spontaneous assembly of the late complement components, C5-C9, into the membrane attack complex. C5b has a transient binding site for C6. The C5b-C6 complex is the foundation upon which the lytic complex is assembled. The C5a anaphylatoxin interacts with C5AR1 and tick complement inhibitor. C5a is also a potent chemokine which stimulates the locomotion of polymorphonuclear leukocytes and directs their migration toward sites of inflammation.
- References
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
ebiomall.com
>
>
>
>
>
>
>
>
>
>
>
>
1 mmol/LEDTA(pH 8.0)
因为含有以上两种物质，所以称为TE。
配制分三步：
1)1 M Tris-HCl (pH 8.0) 50 ml的配制：称取Tris碱6.06 g，加超纯水40 ml溶解，滴加浓HCl约2.1 ml调pH至8.0，定容至50 ml。
2)0.5 M EDTA（pH 8.0）50 ml的配制：称取EDTA-Na2·2H2O 9.306 g，加超纯水35 ml，剧烈搅拌，用约1 g NaOH颗粒调pH至8.0，定容至50 ml。（EDTA二钠盐需加入NaOH将pH调至接近8.0时，才会溶解。）
3)1×TE（10 mM Tris-HCl，pH 8.0；1 mM EDTA，pH 8.0）的配制：
作用：
TE缓冲液是弱碱性,对DNA的碱基有保护性,(DNA在它是的稳定性较好,不易破坏其完整性或产生开环及断裂),包括提取好的DNA也要放在TE缓冲液是保存. 10mMTris-Hcl，pH有7.47.68.0三种。
EDTA调到8.0是为了更好溶解，其他只要调到相应pH就可以。Tris在7－8附近缓冲能力很强，所以加8.0的EDTA下去后，不会改变pH。
TBST中含有Tris-Hcl，NaCl，Tween20这三种物质，是做WESTERNBLOT中常用的一种缓冲液。
TBST缓冲液的配制
1000ml×TBST的配置
先称量NaCl40g，倒入烧杯中，加DDW蒸馏水400ml,再称量NaCl47.6g，倒入刚才的那个烧杯中（PS：由于NaCl的量太多，一次称量不方便，所以分两次称量，且易于溶解）。往烧杯中加入Tris—HCl缓冲液100ml，最后加（吐温20）5ml,转入1000ml容量瓶中，在定容，转移即可。
TBST缓冲液的应用：
1.主要用于免疫组化和原位杂交，酶联免疫等实验中，清洗免疫印。
2.迹膜；
注意事项：
1.TBST缓冲液，PH7.2-7.5；
2.颜色为无色透明液体；
3.为了您的安全和健康，请穿实验服并戴防护手套操作；
1、组成成分：
A、1×TE缓冲液：10mmol/LTris.Cl；1mmol/LEDTA，pH8.0。
B、1×TAE缓冲液：40mmol/LTris-乙酸；2mmol/LEDTA，pH8.0。
C、1×TBE缓冲液：45mmol/LTris-硼酸；1mmol/LEDTA，pH8.0。
2、用途：
A、TE缓冲液：一般用作溶解剂或保持剂，常用于溶解DNA，能稳定储存DNA。
B、TAE缓冲液:生物学中使用最广泛的核酸电泳缓冲液，主要用于DNA的琼脂糖凝胶电泳。
C、TBE缓冲液：生物学中常用的核酸电泳缓冲液，主要用于DNA的琼脂糖凝胶电泳。
3、TAE和TBE缓冲液的选择：
TAE和TBE缓冲液都可以用于DNA的琼脂糖凝胶电泳，两者各有利弊，应根据实际情况选择不同的缓冲液。
4、注意：
TBE浓溶液长时间存放后会形成沉淀物，出现沉淀后应予以废弃。
