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產(chǎn)品訂購信息：
英文名稱  Anti-PHKB 

中文名稱   磷酸化酶β抗體規(guī)格 

別    名  Phosphorylase B; Phosphorylase kinase B; Phosphorylase kinase beta polypeptide; Phosphorylase kinase beta subunit; DKFZp781E15103; FLJ41698; KPBB_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(yīng)   Human, Mouse, Rat, Dog, Pig, Cow, Horse, Rabbit 

產(chǎn)品類型   一抗  

研究領(lǐng)域     細(xì)胞生物 免疫學(xué) 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 124kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human PHKB 

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

磷酸化酶β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 Phosphorylase kinase is a polymer of 16 subunits, four each of alpha, beta, gamma and delta. The alpha subunit includes the skeletal muscle and hepatic isoforms, encoded by two different genes. The beta subunit is the same in both the muscle and hepatic isoforms, encoded by this gene, which is a member of the phosphorylase b kinase regulatory subunit family. The gamma subunit also includes the skeletal muscle and hepatic isoforms, encoded by two different genes. The delta subunit is a calmodulin and can be encoded by three different genes. The gamma subunits contain the active site of the enzyme, whereas the alpha and beta subunits have regulatory functions controlled by phosphorylation. The delta subunit mediates the dependence of the enzyme on calcium concentration. Mutations in this gene cause glycogen storage disease type 9B, also known as phosphorylase kinase deficiency of liver and muscle. Alternatively spliced transcript variants encoding different isoforms have been identified in this gene. Two pseudogenes have been found on chromosomes 14 and 20, respectively.[provided by RefSeq, Feb 2010].

Function : Phosphorylase b kinase catalyzes the phosphorylation of serine in certain substrates, including troponin I. The beta chain acts as a regulatory unit and modulates the activity of the holoenzyme in response to phosphorylation.

Subunit : Hexadecamer of 4 heterotetramers, each composed of alpha, beta, gamma, and delta subunits. Alpha (PHKA1 or PHKA2) and beta (PHKB) are regulatory subunits, gamma (PHKG1 or PHKG2) is the catalytic subunit, and delta is calmodulin.

Subcellular Location : Cell membrane; Lipid-anchor; Cytoplasmic side (Potential).

Although the final Cys may be farnesylated, the terminal tripeptide is probably not removed, and the C-terminus is not methylated.

DISEASE : Defects in PHKB are the cause of glycogen storage disease type 9B (GSD9B) [MIM:261750]; also known as phosphorylase kinase deficiency of liver and muscle (PKD). GSD9B is a metabolic disorder characterized by hepathomegaly, only slightly elevated transaminases and plasma lipids, clinical improvement with increasing age, and remarkably no clinical muscle involvement. Biochemical observations suggest that this mild phenotype is caused by an incomplete holoenzyme that lacks the beta subunit, but that may possess residual activity.

Similarity : Belongs to the phosphorylase b kinase regulatory chain family.

磷酸化酶A是磷酸化酶的活性狀態(tài)，而磷酸化酶B是其無活性狀態(tài)。6-磷酸葡萄糖結(jié)合磷酸化酶后，磷酸化酶由無活性變?yōu)榛罨瘧B(tài)，的加長,之后，又回到原來無活性狀態(tài)，它充當(dāng)了個(gè)橋梁作用。

LIF ELISA Kit 大鼠抑制因子Multi-class antibodies規(guī)格： 48T

Anti-AIMP2/JTV1/FITC 熒光素標(biāo)記抗AIMP2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh FBXL3 FBXL3蛋白抗體 規(guī)格 0.2ml

UK ELISA Kit 大鼠尿激酶 96T

BBS6 英文名稱： 巴德-畢德氏綜合征蛋白BBS6抗體 0.2ml

A4GNT 英文名稱： α1,4-N-乙酰葡糖胺轉(zhuǎn)移酶α4Gn-T抗體 0.2ml

Anti-AIMP2/JTV1/FITC 熒光素標(biāo)記抗AIMP2抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Eukaryotic aspartyl protease(Rice) 天冬氨酸蛋白酶家族蛋白(水稻蛋白的一種)(抗原)Multi-class antibodies規(guī)格： 0.5mg

Anti-CCL25/CCR-9 細(xì)胞表面趨化因子受體9抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh PAX6 轉(zhuǎn)錄因子Pax6抗體 規(guī)格 0.2ml

BAMBI Kit Human 人 BAMBI / NMA ELISA配對抗體 ELISA

TRPV6 英文名稱： 鈣轉(zhuǎn)運(yùn)蛋白1抗體 0.1ml

Claudin 5 英文名稱： 緊密連接蛋白5抗體 0.1ml

Anti-CCL25/CCR-9 細(xì)胞表面趨化因子受體9抗體Multi-class antibodies規(guī)格： 0.2ml

Notch1/MOTC(Neurogenic locus notch homolog protein 1) 跨膜受體蛋白Notch-1抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-AQP3 水通道蛋白-3抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh phospho-ATG4D(Ser467) 磷酸化自噬相關(guān)蛋白4D抗體 規(guī)格 0.1ml

山羊抗人 IgG(H+L)/HRP 0.1ml 美國Jackson公司分裝

VGLL1 英文名稱： 轉(zhuǎn)錄輔助因子退變樣蛋白1抗體 0.2ml

DENND2C 英文名稱： MAP激酶激活死亡域蛋白2C抗體 0.2ml

Anti-AQP3 水通道蛋白-3抗體Multi-class antibodies規(guī)格： 0.1ml

人羊膜上皮細(xì)胞

BRL 大鼠肝細(xì)胞

XPNPEP2 Others Human 人 XPNPEP2 / X-Pro aminopeptidase 2 人細(xì)胞裂解液 (陽性對照)

CL-0126JAR(人胎盤細(xì)胞)5×106cells/瓶×2

人肺微內(nèi)皮細(xì)胞裂解物HPMECL

人正?；|(zhì)永生化細(xì)胞;WPMY-1 人心肌成纖維細(xì)胞完全培養(yǎng)基 100mL

CL-0114Hs 578T(人癌細(xì)胞)5×106cells/瓶×2

KIR2DL4 Others Human 人 KIR2DL4 / CD158D CHO細(xì)胞裂解液 (陽性對照)

人外根殼細(xì)胞裂解物HHORSCL

牛源細(xì)胞 細(xì)胞，HCCLM3細(xì)胞 C3細(xì)胞，小鼠細(xì)胞

人胚腎細(xì)胞(SV40T基因修飾);293T/17

COCH Others Human 人 COCH / Cochlin 人細(xì)胞裂解液 (陽性對照)

磷酸化酶β抗體規(guī)格 人羊膜上皮細(xì)胞

BRL 大鼠肝細(xì)胞

XPNPEP2 Others Human 人 XPNPEP2 / X-Pro aminopeptidase 2 人細(xì)胞裂解液 (陽性對照)

CL-0126JAR(人胎盤細(xì)胞)5×106cells/瓶×2

人肺微內(nèi)皮細(xì)胞裂解物HPMECL

人正常基質(zhì)永生化細(xì)胞;WPMY-1 人心肌成纖維細(xì)胞完全培養(yǎng)基 100mL

抗體的生物素化標(biāo)記實(shí)驗(yàn)要點(diǎn)：

1. 磷酸化酶β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(huì)抑制標(biāo)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(huì)有所不同,選擇不當(dāng)則影響標(biāo)記的效率,應(yīng)先用幾個(gè)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(diǎn)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(shí)可加入去污劑如 Triton x-100, Tween20等；

5.當(dāng)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(diǎn)時(shí),或位于酶的催化位點(diǎn)時(shí),生物素化會(huì)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(tuán),如羰基、氨基、巰基、異咪唑基及苯酚基,也可與糖基共價(jià)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(huì)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(xì)胞的熒光標(biāo)記實(shí)驗(yàn)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(xì)胞可導(dǎo)致高本底。

抗體的鑒定：

1）磷酸化酶β抗體規(guī)格 抗體的效價(jià)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(jià)。不同的抗原制備的抗體,要求的效價(jià)不一。鑒定效價(jià)的方法很多,包括有試管凝集反應(yīng),瓊脂擴(kuò)散試驗(yàn),酶聯(lián)免疫吸附試驗(yàn)等。常用的抗原所制備的抗體一般都有約成的鑒定效價(jià)的方法,以資比較。如制備抗抗體的效價(jià),一般就采用瓊脂擴(kuò)散試驗(yàn)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(qiáng)。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(yàn)測定。以不同濃度抗原和近似抗原分別做競爭抑制曲線,計(jì)算各自的結(jié)合率,求出各自在IC50時(shí)的濃度,并按公式計(jì)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(shí),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(diǎn)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?？贵w親和力的測定對抗體的篩選,確定抗體的用途,驗(yàn)證抗體的均一性等均有重要意義。