抗體的生物素化標記實驗要點：

1. 環(huán)指蛋白12抗體規(guī)格 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之間的分子比按蛋白質表面的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-RNF12 

中文名稱   環(huán)指蛋白12抗體規(guī)格 

別    名  RLIM; RING finger LIM domain-binding protein; Ring finger protein 12; RNF12_HUMAN.

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, Dog, Pig, Cow, Horse, Rabbit, Sheep 

產(chǎn)品類型   一抗  

研究領域    細胞生物 免疫學 表觀遺傳學

蛋白分子量  predicted molecular weight: 69kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human RNF12 

亞    型   IgG

純化方法   affinity purified by Protein A

儲 存 液   Preservative: 15mM Sodium Azide, Constituents: 1% BSA, 0.01M PBS, pH 7.4

環(huán)指蛋白12抗體規(guī)格 產(chǎn)品應用    WB=1:100-500 ELISA=1:500-1000 IHC-P=1:100-500 IHC-F=1:100-500 IF=1:50-200

（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 RLIM, also known as RNF12 (RING finger protein 12) or NY-REN-43, is a 624 amino acid RING-H2 zinc finger protein that is involved in protein ubiquitinylation and subsequent degradation. Expressed in a variety of tissues, RLIM binds to the LIM domain of various proteins and functions as a protein ligase that negatively co-regulates LIM homeodomain (LIM-HD) transcription factors. Through its interaction with Sin3A, a component of the histone deacetylase corepressor complex, RLIM is able to recruit the corepressor complex to LIM-HD proteins, thereby inhibiting LIM-HD transcription. In addition to recruiting the deacetylase complex to LIM-HD proteins, RLIM is able to bind to, ubiquinate and subsequently degrade CLIM proteins, which function as positive co-regulators of LIM-HD transcription factors. RLIM contains one RING-type zinc finger and is implicated in renal cell carcinoma.

Function : E3 ubiquitin-protein ligase. Acts as a negative coregulator for LIM homeodomain transcription factors by mediating the ubiquitination and subsequent degradation of LIM cofactors LDB1 and LDB2 and by mediating the recruitment the SIN3a/histone deacetylase corepressor complex. Ubiquitination and degradation of LIM cofactors LDB1 and LDB2 allows DNA-bound LIM homeodomain transcription factors to interact with other protein partners such as RLIM. Plays a role in telomere length-mediated growth suppression by mediating the ubiquitination and degradation of TERF1. By targeting ZFP42 for degradation, acts as an activator of random inactivation of X chromosome in the embryo, a stochastic process in which one X chromosome is inactivated to minimize sex-related dosage differences of X-encoded genes in somatic cells of female placental mammals.

Subunit : Interacts with LIM/homeobox factors such as LHX3. Interacts with LDB1, LDB2 and SIN3A (By similarity). Interacts with LIMK1 (By similarity). Interacts (via N-terminus) with TERF1. Interacts (via C-terminus) with ESR1.

Subcellular Location : Cytoplasmic and Nuclear

Tissue Specificity : Expressed in many tissues.

Similarity : Belongs to the RNF12 family.

Contains 1 RING-type zinc finger.

Database links : UniProtKB/Swiss-Prot: Q9NVW2.3

抗體的鑒定：

1）環(huán)指蛋白12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,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,抗體分子的結合位點與抗原決定簇之間立體構型的合適度決定的。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。

IFABP(Intestinal Fatty acid binding protein) 小腸型脂肪酸結合蛋白抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-ABP 雄激素結合蛋白抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-CENP-A (Ser7) 磷酸化著絲粒蛋白A 規(guī)格 0.1ml

兔抗人IgG- FC 0.5ml 國產(chǎn)

WNT9A 英文名稱： 信號通路Wnt9a抗體 0.1ml

DAPK1 英文名稱： 凋亡相關蛋白激酶1抗體 0.1ml

Anti-ABP 雄激素結合蛋白抗體Multi-class antibodies規(guī)格： 0.2ml

Anti-PTP-1B/FITC 熒光素標記PTP-1B蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Recomb Protein G/FITC 熒光素(FITC)標記重組蛋白-GMulti-class antibodies規(guī)格： 0.5ml

粘附分子整合素β1抗體 Anti-ITG-β1/CD29 0.1ml

SOX10 英文名稱： 轉錄因子SOX10抗體 0.2ml

FRAS1 英文名稱： 細胞外基質蛋白FRAS1抗體 0.2ml

Rhesus antibody Rh phospho-PPM1A(Ser375) 磷酸化蛋酸酶2C亞型α抗體 規(guī)格 0.1ml

Recomb Protein G/FITC 熒光素(FITC)標記重組蛋白-GMulti-class antibodies規(guī)格： 0.5ml

Phospho-SRF (Ser103) 英文名稱： 磷酸化血清應答因子抗體 0.1ml

EXOC2 英文名稱： 胞吐囊復合體蛋白2抗體 0.2ml

CD45RO抗體 Anti-CD45RO 0.1ml

Anti-SV2A/FITC 熒光素標記突觸泡蛋白2A抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Phospho-Glucocorticoid Receptor (Ser211) 磷酸化糖皮質激素受體抗體 規(guī)格 0.1ml

CDK7/Cyclin H/MAT1 周期素依賴性激酶7抗原Multi-class antibodies規(guī)格： 0.5mg

THPO Protein Human 重組人 Thrombopoietin / THPO / TPO 蛋白 (His 標簽)

MuM-2C 人眼脈絡細胞

CD6 Others Mouse 小鼠 CD6 / TP120 人細胞裂解液 (陽性對照)

hFOB 1.19人SV40轉染成骨細胞 The hFOB 1.19 SV40 was ansfected io osteoblasts DMEM/F12(1:1)+10% FBS+300μg/ml G418

EB病毒轉化的人B淋巴細胞;KMY0927

人胎兒胸腺細胞株;HFT-8810 [HFT8810] 人淋巴內皮細胞完全培養(yǎng)基 100mL

CL-0248ZR-75-30(人癌細胞)5×106cells/瓶×2

OXSR1 Others Human 人 OXSR1 / OSR1 桿狀病毒-昆蟲細胞裂解液 (陽性對照)

人腎近曲小管上皮細胞cDNAHRPTEpiC cDNA

SHZ-88細胞，大鼠癌細胞 人(轉染TK基因的GLC-82細胞),GLC-82-TK細胞 肺泡上皮細胞Many types of cells包裝：5 × 105次方(1ml)

犬胸腺細胞;cf2Th

TNFRSF10D Others Human 人 AILR4 / TNFRSF10D / DCR2 / CD264 人細胞裂解液 (陽性對照)

環(huán)指蛋白12抗體規(guī)格 THPO Protein Human 重組人 Thrombopoietin / THPO / TPO 蛋白 (His 標簽)

MuM-2C 人眼脈絡細胞

CD6 Others Mouse 小鼠 CD6 / TP120 人細胞裂解液 (陽性對照)

hFOB 1.19人SV40轉染成骨細胞 The hFOB 1.19 SV40 was ansfected io osteoblasts DMEM/F12(1:1)+10% FBS+300μg/ml G418

EB病毒轉化的人B淋巴細胞;KMY0927

人胎兒胸腺細胞株;HFT-8810 [HFT8810] 人淋巴內皮細胞完全培養(yǎng)基 100mL