抗體的生物素化標(biāo)記實驗要點：

1. 磷酸化蛋白激酶C(T410)抗體說明書 如在反應(yīng)混合液中有疊氮鈉或游離氨基存在,會抑制標(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-氨基的密度會有所不同,選擇不當(dāng)則影響標(biāo)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(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(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(tuán),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(xì)胞的熒光標(biāo)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(xì)胞可導(dǎo)致高本底。

產(chǎn)品訂購信息：
英文名稱  Anti-phospho-PRKCZ(Thr410) 

中文名稱   磷酸化蛋白激酶C(T410)抗體說明書 

別    名  PRKCZ(phospho T410); PRKCZ(phospho Thr410); p-PRKCZ(T410); rotein Kinase C; AAG6; Aging associated gene 6; MGC129900; MGC129901; MGC41878; MGC57564; PKC B; PKC beta; PKC zeta; PKC2; PKCB; PKCC; PKCD; PKCE; PKCG; PRKACA; PRKC A; PRKC G; PRKCA; PRKCB; PRKCB1; PRKCB2; PRKCD; PRKCE; PRKCG; PRKCZ ; Protein kinase C alpha; Protein kinase C alpha type; Protein Kinase C; Protein kinase C beta 1; Protein kinase C beta 1 polypeptide; Protein kinase C beta; Protein kinase C beta type; Protein kinase C delta; Protein kinase C epsilon; Protein kinase C gamma; Protein kinase C gamma type; Protein kinase C zeta; SCA14; Spinocerebellar ataxia 14.

濃    度   1mg/1ml

規(guī) 格  0.1ml/100μg

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 磷酸化抗體  

研究領(lǐng)域    免疫學(xué) 神經(jīng)生物學(xué) 信號轉(zhuǎn)導(dǎo) 轉(zhuǎn)錄調(diào)節(jié)因子 激酶和磷酸酶

蛋白分子量  predicted molecular weight: 66kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated Synthesised phosphopeptide derived from human PRKCZ around the phosphorylation site of Thr410 

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

磷酸化蛋白激酶C(T410)抗體說明書 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 ICC=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 Protein kinase C (PKC) zeta is a member of the PKC family of serine/threonine kinases which are involved in a variety of cellular processes such as proliferation, differentiation and secretion. Unlike the classical PKC isoenzymes which are calcium-dependent, PKC zeta exhibits a kinase activity which is independent of calcium and diacylglycerol but not of phosphatidylserine. Furthermore, it is insensitive to typical PKC inhibitors and cannot be activated by phorbol ester. Unlike the classical PKC isoenzymes, it has only a single zinc finger module. These structural and biochemical properties indicate that the zeta subspecies is related to, but distinct from other isoenzymes of PKC. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jul 2008].

Function : Calcium- and diacylglycerol-independent serine/threonine-protein kinase that functions in phosphatidylinositol 3-kinase (PI3K) pathway and mitogen-activated protein (MAP) kinase cascade, and is involved in NF-kappa-B activation, mitogenic signaling, cell proliferation, cell polarity, inflammatory response and maintenance of long-term potentiation (LTP). Upon lipopolysaccharide (LPS) treatment in macrophages, or following mitogenic stimuli, functions downstream of PI3K to activate MAP2K1/MEK1-MAPK1/ERK2 signaling cascade independently of RAF1 activation. Required for insulin-dependent activation of AKT3, but may function as an adapter rather than a direct activator. Upon insulin treatment may act as a downstream effector of PI3K and contribute to the activation of translocation of the glucose transporter SLC2A4/GLUT4 and subsequent glucose transport in adipocytes. In EGF-induced cells, binds and activates MAP2K5/MEK5-MAPK7/ERK5 independently of its kinase activity and can activate JUN promoter through MEF2C. Through binding with SQSTM1/p62, functions in interleukin-1 signaling and activation of NF-kappa-B with the specific adapters RIPK1 and TRAF6. Participates in TNF-dependent transactivation of NF-kappa-B by phosphorylating and activating IKBKB kinase, which in turn leads to the degradation of NF-kappa-B inhibitors. In migrating astrocytes, forms a cytoplasmic complex with PARD6A and is recruited by CDC42 to function in the establishment of cell polarity along with the microtubule motor and dynein. In association with FEZ1, stimulates neuronal differentiation in PC12 cells. In inflammatory response, is required for the T-helper 2 (Th2) differentiation process, including interleukins production, efficient activation of JAK1 and the subsequent phosphorylation and nuclear translocation of STAT6. May be involved in development of allergic airway inflammation (asthma), a process dependent on Th2 immune response. In NF-kappa-B-mediated inflammatory response, can relieve the SETD6-dependent repression of NF-kappa-B target genes by phosphorylating the RELA subunit at 'Ser-311'. Is necessary and sufficient for LTP maintenance in hippocampal CA1 pyramidal cells.

Subunit : Forms a ternary complex with SQSTM1 and KCNAB2. Forms another ternary complex with SQSTM1 and GABRR3. Forms a complex with SQSTM1 and MAP2K5 (By similarity). Interacts with PARD6A, PARD6B, PARD6G and SQSTM1. Part of a complex with PARD3, PARD6A or PARD6B or PARD6G and CDC42 or RAC1. Interacts with ADAP1/CENTA1. Forms a ternary complex composed of SQSTM1 and PAWR. Interacts directly with SQSTM1 (Probable). Interacts with IKBKB. Interacts (via the protein kinase domain) with WWC1. Forms a tripartite complex with WWC1 and DDR1, but predominantly in the absence of collagen. Component of the Par polarity complex, composed of at least phosphorylated PRKCZ, PARD3 and TIAM1. Interacts with PDPK1 (via N-terminus region).

Subcellular Location : Cytoplasm. Endosome. Cell junction. Note=In the retina, localizes in the terminals of the rod bipolar cells. Associates with endosomes. Presence of KRIT1, CDH5 and RAP1B is required for its localization to the cell junction.

Post-translational modifications : CDH5 is required for its phosphorylation at Thr-410. Phosphorylated by protein kinase PDPK1; phosphorylation is inhibited by the apoptotic C-terminus cleavage product of PKN2. Phosphorylation at Thr-410 by PI3K activates the kinase.

Similarity : Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. PKC subfamily.

Contains 1 AGC-kinase C-terminal domain.

Contains 1 OPR domain.

Contains 1 phorbol-ester/DAG-type zinc finger.

Contains 1 protein kinase domain.

Database links : UniProtKB/Swiss-Prot: Q05513.4

抗體的鑒定：

1）磷酸化蛋白激酶C(T410)抗體說明書 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(kuò)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(kuò)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(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(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。

Anti-Apollon/FITC 熒光素標(biāo)記Apollon凋亡抑制蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-Phospho-FHIT (Tyr114) /FITC 熒光素標(biāo)記抗磷酸化脆性組氨酸三聯(lián)體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh Collagen IV 抗IV型膠原抗體 規(guī)格 0.1ml

SPAG5/map126 相關(guān)抗原5(多肽抗原) 0.5mg

phospho-JMJD2B(Thr305) 英文名稱： 磷酸化組蛋白去甲基化酶JMJD2B抗體 0.1ml

Rhesus antibody Rh TTC3/RNF105 環(huán)指蛋白105抗體 規(guī)格 0.2ml

Anti-Phospho-FHIT (Tyr114) /FITC 熒光素標(biāo)記抗磷酸化脆性組氨酸三聯(lián)體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti- Beta-lactoglobulin/FITC 熒光素標(biāo)記β-乳球蛋白抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Anti-SNAP-25 /FITC 熒光素標(biāo)記突觸相關(guān)蛋白25抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh AKT/PKB 蛋白激酶B 規(guī)格 0.1ml

Rabbit Anti-horse IgG/PE-CY5 PE-CY5標(biāo)記的兔抗馬IgG 0.1ml

Gab1 英文名稱： 接頭蛋白Gab 1抗體 0.1ml

Rhesus antibody Rh Rabbit Anti-Guinea pig IgG/Alexa Fluor 350 Alexa Fluor 350標(biāo)記的兔抗豚鼠IgG 規(guī)格 0.1ml

Anti-SNAP-25 /FITC 熒光素標(biāo)記突觸相關(guān)蛋白25抗體IgGMulti-class antibodies規(guī)格： 0.2ml

PIIINP 大鼠III型前膠原肽(N端)Multi-class antibodies規(guī)格： 48T

Anti-DKK1 蛋白DKK1抗體Multi-class antibodies規(guī)格： 0.2ml

Rhesus antibody Rh NDE1 核分布基因E同源蛋白1抗體 規(guī)格 0.2ml

Human apoprotein A1,apo-A1 ELISA Kit 人載脂蛋白A1 96T

SLC1A5 英文名稱： 溶質(zhì)攜帶物家族-1抗體 0.2ml

CRF 英文名稱： 促腎上腺皮質(zhì)激素釋放因子抗體 0.1ml

Anti-DKK1 蛋白DKK1抗體Multi-class antibodies規(guī)格： 0.2ml

GlucoseAsparagineMedium

DMEM(低糖)，干粉 "含1000mg/l葡萄糖，L-谷氨酰胺， incubation media DMEM(低糖)，干粉 "含1000mg/l葡萄糖，L-谷氨酰胺，

酵米面假單胞菌(酵米面黃桿菌) 支/瓶

CIN-(9cm)用于分離小腸耶爾森氏菌

RVSalmonellaEnrichmentMedium

抗生素檢定培養(yǎng)基H)(05藥典)250g用于磺芐青霉素效價測定

馬鈴薯瓊脂 100(g) incubation media 馬鈴薯瓊脂 100(g)

磷酸鹽葡萄糖蛋白胨水培養(yǎng)基 Phosphate Glucose Peptone Water 250克 用于甲基紅試驗

菌種培養(yǎng)基250用于四環(huán)素檢定中蠟樣芽孢桿菌的培養(yǎng)(SN標(biāo)準(zhǔn))incubationmedia菌種培養(yǎng)基250用于四環(huán)素檢定中蠟樣芽孢桿菌的培養(yǎng)(SN標(biāo)準(zhǔn))

我妻氏培養(yǎng)基基礎(chǔ) 250g 用于副溶血性弧菌神奈川現(xiàn)象試驗(GB標(biāo)準(zhǔn)、SN)

磷酸化蛋白激酶C(T410)抗體說明書 亮綠乳糖液體培養(yǎng)基（BGB）  規(guī)格：  250g  用途：  用于多管發(fā)酵法測定飲用礦泉水中大腸菌群的確證試驗(GB/T8538-2008)。

品紅亞培養(yǎng)基（遠(yuǎn)藤瓊脂培養(yǎng)基）  規(guī)格：  250g  用途：  用于水中大腸菌群的分離或確證試驗。 (GB/T8538-2008和GB5750.12—2006)。

乳糖膽鹽發(fā)酵培養(yǎng)基  規(guī)格：  250g  用途：  用于多管發(fā)酵法測定食品、藥品、純凈水、礦泉水和一次性使用衛(wèi)生用品中大腸菌群、糞大腸菌群和大...

大腸菌群顯色培養(yǎng)基  規(guī)格：  1000ml  用途：  用于大腸菌群的快速檢測和計數(shù)。