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抗體的生物素化標記實驗要點：

1. 轉(zhuǎn)錄中介因子Tif1γ抗體規(guī)格 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會抑制標記反應(yīng)。因此,蛋白質(zhì)在反應(yīng)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(yīng)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(yīng)后,應(yīng)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-TIF1 gamma/Trim33 

中文名稱   轉(zhuǎn)錄中介因子Tif1γ抗體規(guī)格 

別    名  RFG7; Rfg7 protein; TF1G; TIF1-gamma; TIF1G; TIF 1G; TIF1GAMMA; TIFGAMMA; Transcription intermediary factor 1-gamma; Transcriptional intermediary factor 1 gamma； Trim 33; Tripartite motif containing 33; Tripartite motif containing 33 protein; tripartite motif-containing 33; Tripartite motif-containing protein 33; E3 ubiquitin-protein ligase TRIM33; Ectodermin homolog; Protein Rfg7; PTC7 antibody; Ret fused gene 7; RET-fused gene 7 protein; RFG7; KIAA1113.

濃    度   1mg/1ml

規(guī) 格  0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Horse 

產(chǎn)品類型   一抗  

研究領(lǐng)域     免疫學 信號轉(zhuǎn)導 轉(zhuǎn)錄調(diào)節(jié)因子

蛋白分子量  predicted molecular weight: 124kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human TIF1 gamma(987-1048aa) 

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

轉(zhuǎn)錄中介因子Tif1γ抗體規(guī)格 產(chǎn)品應(yīng)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 TIF1 gamma is a member of the Transcriptional Intermediary Factor 1 family, which serve as coactivators and corepressors. Its structure is similar to TIF1 alpha and TIF1 beta, exhibiting multiple domains (RING finger, coiled coil, B boxes, PHD/TTC, and bromodomain). TIF1 gamma functions in cell differentiation and development, playing a role in differentiation of hematopoietic cells.

Function : Acts as an E3 ubiquitin-protein ligase. Promotes SMAD4 ubiquitination, nuclear exclusion and degradation via the ubiquitin proteasome pathway. According to PubMed:16751102, does not promote a decrease in the level of endogenous SMAD4. May act as a transcriptional repressor. Inhibits the transcriptional response to TGF-beta/BMP signaling cascade. Plays a role in the control of cell proliferation. Its association with SMAD2 and SMAD3 stimulates erythroid differentiation of hematopoietic stem/progenitor (By similarity). Monoubiquitinates SMAD4 and acts as an inhibitor of SMAD4-dependent TGF-beta/BMP signaling cascade (Monoubiquitination of SMAD4 hampers its ability to form a stable complex with activated SMAD2/3 resulting in inhibition of TGF-beta/BMP signaling cascade).

Subunit : Homooligomer and heterooligomer with TRIM24 and TRIM28 family members. Interacts with SMAD4 in unstimulated cells. Found in a complex with SMAD2 and SMAD3 upon addition of TGF-beta. Interacts with SMAD2 and SMAD3. Interacts with SMAD4 under basal and induced conditions and, upon TGF-beta signaling, with activated SMAD2. Forms a ternary complex with SMAD4 and SMAD2 upon TGF-beta signaling.

Subcellular Location : Nucleus. Note=In discrete nuclear dots resembling nuclear bodies (By similarity).

Tissue Specificity : Expressed in stem cells at the bottom of the crypts of the colon (at protein level). Expressed in colon adenomas and adenocarcinomas (at protein level). Expressed in brain, lung, liver, spleen, thymus, prostate, kidney, testis, heart, placenta, pancreas, small intestine, ovary, colon, skeletal muscle and hematopoietic progenitors.

DISEASE : Defects in TRIM33 are a cause of thyroid papillary carcinoma (TPC) [MIM:188550]. TPC is a common tumor of the thyroid that typically arises as an irregular, solid or cystic mass from otherwise normal thyroid tissue. Papillary carcinomas are malignant neoplasm characterized by the formation of numerous, irregular, finger-like projections of fibrous stroma that is covered with a surface layer of neoplastic epithelial cells. Note=A chromosomal aberration involving TRIM33 is found in thyroid papillary carcinomas. Translocation t(1;10)(p13;q11) with RET. The translocation generates the TRIM33/RET (PTC7) oncogene.

Similarity : Belongs to the TRIM/RBCC family.

Contains 2 B box-type zinc fingers.

Contains 1 bromo domain.

Contains 1 PHD-type zinc finger.

Contains 1 RING-type zinc finger.

抗體的鑒定：

1）轉(zhuǎn)錄中介因子Tif1γ抗體規(guī)格 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(yīng),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(yīng)抗原或近似抗原物質(zhì)的識別能力。抗體的特異性高,它的識別能力就強。衡量特異性通常以交叉反應(yīng)率來表示。交叉反應(yīng)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(yīng)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(yīng)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?？贵w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。

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Rhesus antibody Rh p53BP1/53BP1 p53結(jié)合蛋白1抗體 規(guī)格 0.2ml

IL13RA1 Kit Mouse 小鼠 IL-13RA1 / CD213a1 ELISA配對抗體 ELISA

TSPAN10 英文名稱： 四分子交聯(lián)體10抗體(四旋蛋白) 0.2ml

Cathelicidin 英文名稱： 抗菌肽CAMP抗體 0.1ml

Anti-CD44v6 CD44V6抗體Multi-class antibodies規(guī)格： 0.1ml

NAP-2/CXCL7 ELISA Kit 大鼠中性粒細胞趨化蛋白2Multi-class antibodies規(guī)格： 48T

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Anti-ApoB/FITC 熒光素標記載脂蛋白B抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Visfatin-2(CT)/FITC 熒光素標記內(nèi)脂素/內(nèi)臟脂肪素(C端抗體)IgGMulti-class antibodies規(guī)格： 0.2ml

CK14/17/42/10 peptide 細胞角蛋白14抗原Multi-class antibodies規(guī)格： 0.5mg

細胞周期調(diào)控因子34 Anti-Cdc34 0.1ml

SHARPIN 英文名稱： 線性泛素鏈相關(guān)蛋白SHARPIN抗體 0.2ml

Epsin 1 英文名稱： 內(nèi)吞作用輔助蛋白EPN1抗體 0.2ml

Rhesus antibody Rh phospho-GAB2 (Tyr643) 磷酸化接頭蛋白Gab2抗體 規(guī)格 0.1ml

CK14/17/42/10 peptide 細胞角蛋白14抗原Multi-class antibodies規(guī)格： 0.5mg

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