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轉(zhuǎn)錄中介因子Tif1β抗體規(guī)格
  • 品牌:上海莼試
  • 產(chǎn)地:進口、國產(chǎn)
  • 貨號:CS11914
  • 發(fā)布日期: 2019-01-25
  • 更新日期: 2025-04-22
產(chǎn)品詳請
產(chǎn)地 進口、國產(chǎn)
品牌 上海莼試
保存條件 Store at -20 °C
貨號 CS11914
應(yīng)用范圍 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
CAS編號
抗體名 Anti-TIF1 beta/KAP1/TRIM28
克隆性
靶點 詳見說明書
適應(yīng)物種 詳見說明書
形態(tài) 詳見說明書
宿主 詳見說明書
亞型 IgG
標識物 詳見說明書
濃度 1mg/1ml%
免疫原 KLH conjugated synthetic peptide derived from human TIF1 beta

抗體的生物素化標記實驗要點:

1. 轉(zhuǎn)錄中介因子Tif1β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.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(dǎo)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(dǎo)致高本底。

產(chǎn)品訂購信息:
英文名稱  Anti-TIF1 beta/KAP1/TRIM28 

中文名稱   轉(zhuǎn)錄中介因子Tif1β抗體規(guī)格 

      E3 SUMO protein ligase TRIM28; E3 SUMO-protein ligase TRIM28; FLJ29029; KAP 1; KAP-1; KRAB associated protein 1; KRAB interacting protein 1; KRAB-associated protein 1; KRAB-interacting protein 1; KRIP 1; KRIP-1; KRIP1; Nuclear corepressor KAP 1; Nuclear corepressor KAP-1; RING finger protein 96; RNF96; TF1B; TIF1 beta; TIF1-beta; TIF1B; TIF1B_HUMAN; Transcription intermediary factor 1 beta; Transcription intermediary factor 1-beta; TRIM28; Tripartite motif containing 28; tripartite motif containing protein 28; Tripartite motif-containing protein 28.


       1mg/1ml

規(guī)   0.1ml/100μg 0.2ml/200μg

抗體來源   Rabbit

克隆類型   polyclonal

交叉反應(yīng)   Human, Mouse, Rat, Dog, Cow, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域     細胞生物 免疫學(xué) 染色質(zhì)和核信號 信號轉(zhuǎn)導(dǎo) 細胞凋亡 轉(zhuǎn)錄調(diào)節(jié)因子 激酶和磷酸酶 結(jié)合蛋白 表觀遺傳學(xué) 泛素

蛋白分子量  predicted molecular weight: 88kDa

       Lyophilized or Liquid

 KLH conjugated synthetic peptide derived from human TIF1 beta 

       IgG

純化方法   affinity purified by Protein A

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

轉(zhuǎn)錄中介因子Tif1β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(fù)) 

 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 The KRAB (Kruppel Associated Box) domain is minimally about 45 amino acids in length and is a transcriptional repression domain found in numerous transcription factors. Over 220 KRAB zinc finger protein (KRAB ZFP) genes have been identified in the human genome. These proteins functionally repress transcription via specific interactions with KAP1 (KRAB Associated Protein 1). KAP1 is an 835 amino acid polypeptide that contains a RING finger, B boxes, and a PHD finger. KAP1 has been shown to form complexes with KRAB domain transcription factors and increase the efficiency with which they mediate repression. KAP1 has also been shown to directly interact with HP1 (heterochromatin protein 1) and KRAZ1 (Kruppel associated box containing zinc finger protein 1). KAP1 directly targets KRAZ1 to the foci of centromeric heterochromatin containing HP1alpha, helping to regulate transcriptional repression. Recent studies have shown that KAP1 mutants with the ability to bind KRAB but unable to bind HP1 leads to random distribution of KRAZ1 and strong transcriptional activation.

Function : Nuclear corepressor for KRAB domain-containing zinc finger proteins (KRAB-ZFPs). Mediates gene silencing by recruiting CHD3, a subunit of the nucleosome remodeling and deacetylation (NuRD) complex, and SETDB1 (which specifically methylates histone H3 at 'Lys-9' (H3K9me)) to the promoter regions of KRAB target genes. Enhances transcriptional repression by coordinating the increase in H3K9me, the decrease in histone H3 'Lys-9 and 'Lys-14' acetylation (H3K9ac and H3K14ac, respectively) and the disposition of HP1 proteins to silence gene expression. Recruitment of SETDB1 induces heterochromatinization. May play a role as a coactivator for CEBPB and NR3C1 in the transcriptional activation of ORM1. Also corepressor for ERBB4. Inhibits E2F1 activity by stimulating E2F1-HDAC1 complex formation and inhibiting E2F1 acetylation. May serve as a partial backup to prevent E2F1-mediated apoptosis in the absence of RB1. Important regulator of CDKN1A/p21(CIP1). Has E3 SUMO-protein ligase activity toward itself via its PHD-type zinc finger. Also specifically sumoylates IRF7, thereby inhibiting its transactivation activity. Ubiquitinates p53/TP53 leading to its proteosomal degradation; the function is enhanced by MAGEC2 and MAGEA2, and possibly MAGEA3 and MAGEA6.

Subunit : Oligomer; the RBCC domain homotrimerizes and interacts with one molecule of KRAB to form the KRAB-KAP1 corepressor complex. Binding to a KRAB domain is an absolute requirement for silencing gene expression. Interacts with CEBPB and NR3C1 (By similarity). Interacts with a number of KRAB-ZFP proteins including ZNF10, ZFP53, ZFP68, ZNF382 and ZNF256. Interacts with NCOR1, NR3C1 and CHD3. Interacts with CEBPB (via the RING-type and PHD-type zinc fingers). Component of a ternary complex that includes TRIM28, a HP1 protein (CBX1, CBX3 OR CBX5), a KRAB domain-containing protein, and DNA. Interacts with CBX5 (via the PxVxL motif); the interaction occurs in interphase nuclei and competes for binding POGZ. Interacts with POGZ; the interaction competes for interaction with CBX5. Interacts with SETDB1; the interaction is enhanced by KAP1 sumoylation, stimulates SETB1 histone methyltransferase activity and gene silencing. Interacts (via the PHD-type zinc finger) with UBE2I; the interaction is required for sumoylation and repressor activity. Component of the TRIM28/KAP1-ERBB4-MDM2 complex involved in connecting growth factor and DNA damage responses. Interacts directly with ERBB4; the interaction represses ERBB4-mediated transcription activity. Interacts with MDM2; the interaction contributes to p53/TP53 inactivation. Component of the TRIM28/KAP1-MDM2-p53/TP53; involved in regulating p53/TP53 stabilization and activity. Interacts (via the leucine zipper alpha helical coiled-coil) with E2F1 (central region); the interaction inhibits E2F1 acetylation and transcriptional activity. Interacts with PPP1CA; the interaction dephosphorylates TRIM28 at Ser-824 and forms a complex at the p21 promoter site. Interacts with PPP1CB; the interaction is weak but is increased on dephosphorylation at Ser-824. Interacts with FES/FPS. Interacts with SMARCAD1. Interacts with, and sumoylates IRF7. Interacts with MAGEC2. Part of a complex composed of TRIM28, HDAC1, HDAC2 and EHMT2.

Subcellular Location : Nucleus. Note=Associated with centromeric heterochromatin during cell differentiation through CBX1 (By similarity).

Tissue Specificity : Expressed in all tissues tested including spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral blood leukocytes.

Post-translational modifications : ATM-induced phosphorylation on Ser-824 represses sumoylation leading to the de-repression of expression of a subset of genes involved in cell cycle control and apoptosis in response to genotoxic stress. Dephosphorylation by the phosphatases, PPP1CA and PP1CB forms, allows sumoylation and expression of TRIM28 target genes.

Sumoylation/desumoylation events regulate TRIM28-mediated transcriptional repression. Sumoylation is required for interaction with CHD3 and SETDB1 and the corepressor activity. Represses and is repressed by Ser-824 phosphorylation. Enhances the TRIM28 corepressor activity, inhibiting transcriptional activity of a number of genes including GADD45A and CDKN1A/p21. Lys-554, Lys-779 and Lys-804 are the major sites of sumoylation. In response to Dox-induced DNA damage, enhanced phosphorylation on Ser-824 prevents sumoylation and allows de-repression of CDKN1A/p21.

Auto-ubiquitinated; enhanced by MAGEA2 and MAGEC2.

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.

Database links : UniProtKB/Swiss-Prot: Q13263.5

抗體的鑒定:

1轉(zhuǎn)錄中介因子Tif1β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??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(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/,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(fù)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。

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