抗體的生物素化標記實驗要點：

1. 基底膜相關(guān)軟骨素蛋白多糖抗體品牌 如在反應混合液中有疊氮鈉或游離氨基存在,會抑制標記反應。因此,蛋白質(zhì)在反應前要對 0.1mol/L碳酸氫鈉緩沖液或0.5mol/L硼酸緩沖液充分透析；

2.所用的NHSB及待生物素化蛋白質(zhì)之間的分子比按蛋白質(zhì)表面的ε-氨基的密度會有所不同,選擇不當則影響標記的效率,應先用幾個不同的分子比來篩選最適條件；

3.用NHSB量過量也是不利的,抗原的結(jié)合位點可能因此被封閉,導致抗體失活；

4.由于抗體的氨基不易接近可能造成生物素化不足,此時可加入去污劑如 Triton x-100, Tween20等；

5.當游離ε-氨基(賴氨酸殘基的氨基)存在于抗體的抗原結(jié)合位點時,或位于酶的催化位點時,生物素化會降低或損傷抗體蛋白的結(jié)合力或活性；

6.生物素還可能與不同的功能基團,如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價結(jié)合；

7.交聯(lián)反應后,應充分透析,否則,殘余的生物素會對生物素化抗體與親和素的結(jié)合產(chǎn)生競爭作用；

8.在細胞的熒光標記實驗中,中和親和素的本底低,但由于鏈霉親和素含有少量正電荷,故對某些細胞可導致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-SMC3/CSPG6 

中文名稱   基底膜相關(guān)軟骨素蛋白多糖抗體品牌 

別    名  BAM; Bamacan; Basement membrane associated chondroitin proteoglycan; Basement membrane-associated chondroitin proteoglycan; BMH; CDLS3; chondroitin sulfate proteoglycan 6 (bamacan); Chondroitin sulfate proteoglycan 6; Chromosome associated polypeptide; Chromosome-associated polypeptide; CSPG 6; CSPG6; hCAP; SMC 3; SMC protein 3; SMC-3; smc3; SMC3_HUMAN; SMC3L1; Structural maintenance of chromosome 3; Structural maintenance of chromosomes 3; Structural maintenance of chromosomes protein 3.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應   Human, Mouse, Rat, Chicken, Dog, Cow, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域     細胞生物 免疫學 染色質(zhì)和核信號 細胞周期蛋白 結(jié)合蛋白 細胞分化 表觀遺傳學

蛋白分子量  predicted molecular weight: 142kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human SMC3/CSPG6 

亞    型   IgG

純化方法   affinity purified by Protein A

儲 存 液   0.01M PBS, pH 7.4 with 10 mg/ml BSA and 0.1% Sodium azide

基底膜相關(guān)軟骨素蛋白多糖抗體品牌 產(chǎn)品應用    WB=1:100-500 ELISA=1:500-1000 IP=1:20-100 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:100-500

（石蠟切片需做抗原修復） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存條件  Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. The lyophilized antibody is stable at room temperature for at least one month and for greater than a year when kept at -20°C. When reconstituted in sterile pH 7.4 0.01M PBS or diluent of antibody the antibody is stable for at least two weeks at 2-4 °C. 

Important Note  This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications. 

產(chǎn)品介紹 Central component of cohesin, a complex required for chromosome cohesion during the cell cycle. The cohesin complex may form a large proteinaceous ring within which sister chromatids can be trapped. At anaphase, the complex is cleaved and dissociates from chromatin, allowing sister chromatids to segregate. Cohesion is coupled to DNA replication and is involved in DNA repair. The cohesin complex plays also an important role in spindle pole assembly during mitosis and in chromosomes movement.

Function : Central component of cohesin, a complex required for chromosome cohesion during the cell cycle. The cohesin complex may form a large proteinaceous ring within which sister chromatids can be trapped. At anaphase, the complex is cleaved and dissociates from chromatin, allowing sister chromatids to segregate. Cohesion is coupled to DNA replication and is involved in DNA repair. The cohesin complex plays also an important role in spindle pole assembly during mitosis and in chromosomes movement.

Subunit : Interacts with MXI1, MXD3 and MXD4. Interacts with SYCP2. Found in a complex with SMC1A, CDCA5 and RAD21, PDS5A/APRIN and PDS5B/SCC-112 (By similarity). Forms a heterodimer with SMC1A or SMC1B in cohesin complexes. Cohesin complexes are composed of the SMC1 (SMC1A or SMC1B) and SMC3 heterodimer attached via their hinge domain, RAD21 which link them, and one STAG protein (STAG1, STAG2 or STAG3), which interacts with RAD21. Also found in meiosis-specific cohesin complexes. Interacts with NUMA1, and forms a ternary complex with KIF3B and KIFAP3, suggesting a function in tethering the chromosomes to the spindle pole and in chromosome movement. Interacts with PDS5A and WAPAL; regulated by SMC3 acetylation. Interacts with RPGR (By similarity).

Subcellular Location : Nucleus. Chromosome. Chromosome, centromere. Note=Associates with chromatin. Before prophase it is scattered along chromosome arms. During prophase, most of cohesin complexes dissociate from chromatin probably because of phosphorylation by PLK, except at centromeres, where cohesin complexes remain. At anaphase, the RAD21 subunit of the cohesin complex is cleaved, leading to the dissociation of the complex from chromosomes, allowing chromosome separation. The phosphorylated form at Ser-1083 is preferentially associated with unsynapsed chromosomal regions (By similarity).

Post-translational modifications : Phosphorylated at Ser-1083 in a SPO11-dependent manner (By similarity).

Acetylation at Lys-105 and Lys-106 by ESCO1 is important for genome stability and S phase sister chromatid cohesion. Regulated by DSCC1, it is required for processive DNA synthesis, coupling sister chromatid cohesion establishment during S phase to DNA replication. Deacetylation by HDAC8, regulates release of the cohesin complex from chromatin.

DISEASE : Cornelia de Lange syndrome 3 (CDLS3) [MIM:610759]: A form of Cornelia de Lange syndrome, a clinically heterogeneous developmental disorder associated with malformations affecting multiple systems. Characterized by facial dysmorphisms, abnormal hands and feet, growth delay, cognitive retardation, hirsutism, gastroesophageal dysfunction and cardiac, ophthalmologic and genitourinary anomalies. Cornelia de Lange syndrome type 3 is a mild form with absence of major structural anomalies. The phenotype in some instances approaches that of apparently non-syndromic mental retardation. Note=The disease is caused by mutations affecting the gene represented in this entry.

Similarity : Belongs to the SMC family. SMC3 subfamily.

Database links : UniProtKB/Swiss-Prot: Q9UQE7.2

抗體的鑒定：

1）基底膜相關(guān)軟骨素蛋白多糖抗體品牌 抗體的效價鑒定：不管是用于診斷還是用于,制備抗體的目的都是要求較高效價。不同的抗原制備的抗體,要求的效價不一。鑒定效價的方法很多,包括有試管凝集反應,瓊脂擴散試驗,酶聯(lián)免疫吸附試驗等。常用的抗原所制備的抗體一般都有約成的鑒定效價的方法,以資比較。如制備抗抗體的效價,一般就采用瓊脂擴散試驗來鑒定。

2）抗體的特異性鑒定：抗體的特異性是指與相應抗原或近似抗原物質(zhì)的識別能力?？贵w的特異性高,它的識別能力就強。衡量特異性通常以交叉反應率來表示。交叉反應率可用競爭抑制試驗測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計算各自的結(jié)合率,求出各自在IC50時的濃度,并按公式計算交叉反應率。 

如果所用抗原濃度IC50濃度為pg/管,而一些近似抗原物質(zhì)的IC50濃度幾乎是無窮大時,表示這一抗血清與其他抗原物質(zhì)的交叉反應率近似為0,即該血清的特異性較好。

3）抗體親和力：是指抗體和抗原結(jié)合的牢固程度。親和力的高低是由抗原分子的大小,抗體分子的結(jié)合位點與抗原決定簇之間立體構(gòu)型的合適度決定的。有助于維持抗原抗體復合物穩(wěn)定的分子間力有氫鍵,疏水鍵,側(cè)鏈相反電荷基因的庫侖力,范德華力和空間斥力。親和力常以親和常數(shù)K表示,K的單位是L/mol。抗體親和力的測定對抗體的篩選,確定抗體的用途,驗證抗體的均一性等均有重要意義。

Anti-TRF2/FITC 熒光素標記端粒結(jié)合蛋白2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Alpha-Synuclein 核突觸蛋白(抗原)Multi-class antibodies規(guī)格： 0.5mg

促甲狀腺素受體抗體 Anti-TSHR 0.1ml

C14orf174 英文名稱： 14號染色體開放閱讀框174 0.2ml

EBP50 英文名稱： 細胞骨架連接質(zhì)膜蛋白抗體 0.1ml

Rhesus antibody Rh phospho-IRAK4(Thr345) 磷酸化白介素-1受體相關(guān)激酶4抗體 規(guī)格 0.1ml

Alpha-Synuclein 核突觸蛋白(抗原)Multi-class antibodies規(guī)格： 0.5mg

Anti-Phospho-Rb (Ser780)/FITC 熒光素標記磷酸化視網(wǎng)膜母細胞瘤相關(guān)蛋白1抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rabbit Anti-Guinea IgG Whole serum 兔抗豚鼠IgG抗血清Multi-class antibodies規(guī)格： 1ml

KB抑制蛋白抗體 Anti-IKB β 0.2ml

Rabbit anti-Bovine IgG whole serum 兔抗IgG抗血清 1ml

phospho-FANCA (Ser1149) 英文名稱： 磷酸化范可尼組蛋白A抗體 0.1ml

Rhesus antibody Rh phospho-PTPRA(Tyr798) 磷酸化酪氨酸磷酸酶α抗體 規(guī)格 0.1ml

Rabbit Anti-Guinea IgG Whole serum 兔抗豚鼠IgG抗血清Multi-class antibodies規(guī)格： 1ml

Integrin Alpha3 整合素α3抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-ADH/AVP 抗激素/升壓素抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-cdc25A (Ser292) 磷酸化細胞分裂周期蛋白25抗體 規(guī)格 0.1ml

核快紅染色液-淺紅色 6ml Sigma原料

VCP 英文名稱： 含纈酪肽蛋白抗體 0.1ml

phospho-Dynamin 1 (Ser774) 英文名稱： 磷酸化酶動力蛋白1抗體 0.1ml

Anti-ADH/AVP 抗激素/升壓素抗體Multi-class antibodies規(guī)格： 0.2ml

IFNG Protein Mouse 重組小鼠 IFNG / Ierferon Gamma 蛋白 (His 標簽)

大鼠神經(jīng)膠質(zhì)細胞完全培養(yǎng)基 100mL

GDNF Others Human 人 GDNF 人細胞裂解液 (陽性對照)

MKN-45人細胞 Human gasic cancer cells MKN-45 1640+10% FBS

MGC80-3(MGC-803)人細胞

NCI-H157(人非小細胞) 5×106cells/瓶×2 SRSV/3T3(SRSV轉(zhuǎn)化的3T3細胞)

CM-R095大鼠成年表皮角質(zhì)形成層細胞完全培養(yǎng)基100mL

NP Others H7N9 甲型 H7N9 (A/Shanghai/1/2013) Nucleocapsid / NP 桿狀病毒-昆蟲細胞裂解液 (陽性對照)

小鼠胚胎成纖維細胞-處理MEF-mt

非洲綠猴腎細胞;Vero E6 人類惡性，A375細胞 LS 174T(腺癌細胞)

EB病毒轉(zhuǎn)化的人B淋巴細胞;KMY0922

DMBT1 Others Human 人 DMBT1 / GP340 人細胞裂解液 (陽性對照)

基底膜相關(guān)軟骨素蛋白多糖抗體品牌 IFNG Protein Mouse 重組小鼠 IFNG / Ierferon Gamma 蛋白 (His 標簽)

大鼠神經(jīng)膠質(zhì)細胞完全培養(yǎng)基 100mL

GDNF Others Human 人 GDNF 人細胞裂解液 (陽性對照)

MKN-45人細胞 Human gasic cancer cells MKN-45 1640+10% FBS

MGC80-3(MGC-803)人細胞

NCI-H157(人非小細胞) 5×106cells/瓶×2 SRSV/3T3(SRSV轉(zhuǎn)化的3T3細胞)