抗體的生物素化標記實驗要點：

1. AFG3樣蛋白2/脊髓小腦蛋白28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(zhì)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(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-SCA28/AFG3L2 

中文名稱   AFG3樣蛋白2/脊髓小腦蛋白28抗體規(guī)格 

別    名  AFG3 (ATPase family gene 3, yeast) like 2; AFG3 ATPase family gene 3 like 2 (yeast); AFG3 ATPase family gene 3 like 2; AFG3 like protein 2; ATPase family gene 3 like 2; ATPase family gene 3 yeast; FLJ25993; Paraplegin like protein; SCA28; Spinocerebellar ataxia 28; AFG32_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, Chicken, Dog, Pig, Cow, Horse, Sheep 

產(chǎn)品類型   一抗  

研究領域     細胞生物 神經(jīng)生物學 信號轉(zhuǎn)導 泛素

蛋白分子量  predicted molecular weight: 89kDa

性    狀   Lyophilized or Liquid

免 疫 原 KLH conjugated synthetic peptide derived from human AFG3L2 (531-600aa) 

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

AFG3樣蛋白2/脊髓小腦蛋白28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 ICC=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 AFG3L2 is a multi-pass membrane metalloprotease that contains one AAA (ATPase associated with diverse cellular activities) domain, a zinc-dependent binding motif, an RNA-binding region and an ATP/GTP binding site. Localizing to the mitochondrial membrane, AFG3L2 is ubiquitously expressed with highest expression levels in skeletal muscle and heart. AFG3L2 shares 69% similarity with the yeast Afg3 protein and 49% similarity with Paraplegin, a protein of mitochondria that is thought to be involved in signal transduction and chaperone-like activities. In mitochondria, AFG3L2 forms a complex with Paraplegin that is believed to regulate essential protein quality control. Mutations in the gene encoding either one of these proteins can result in hereditary spastic paraplegia, a degenerative spinal cord disorder that is characterized by muscle spasms, stiffness in the legs and, in some cases, incontinence.

Function : AFG3L2 is a protein localized in mitochondria and closely related to paraplegin. The paraplegin gene is responsible for an autosomal recessive form of hereditary spastic paraplegia. AFG3L2 is a candidate gene for other hereditary spastic paraplegias or neurodegenerative disorders and is a putative ATP dependent protease

Subunit : Homooligomer. Interacts with SPG7; the interaction is required for the efficient assembly of mitochondrial complex I.

Subcellular Location : Mitochondrial membrane; multipass membrane protein

Tissue Specificity : Ubiquitous. Highly expressed in the cerebellar Purkinje cells.

DISEASE : Defects in AFG3L2 are the cause of spinocerebellar ataxia type 28 (SCA28) [MIM:610246]. It is a clinically and genetically heterogeneous group of cerebellar disorders. Patients show progressive incoordination of gait and often poor coordination of hands, speech and eye movements, due to degeneration of the cerebellum with variable involvement of the brainstem and spinal cord. SCA28 is an autosomal dominant cerebellar ataxia (ADCA) with a slow progressive course and no evidence of sensory involvement or cognitive impairment.

Defects in AFG3L2 are the cause of spastic ataxia autosomal recessive type 5 (SPAX5) [MIM:614487]. A neurodegenerative disorder characterized by early onset spasticity, peripheral neuropathy, ptosis, oculomotor apraxia, dystonia, cerebellar atrophy, and progressive myoclonic epilepsy.

Similarity : In the N-terminal section; belongs to the AAA ATPase family.

In the C-terminal section; belongs to the peptidase M41 family.

Database links : UniProtKB/Swiss-Prot: Q9Y4W6.2

抗體的鑒定：

1）AFG3樣蛋白2/脊髓小腦蛋白28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(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(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(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。

Anti-STAT3 /FITC 熒光素標記信號轉(zhuǎn)導和轉(zhuǎn)錄激活因子3抗體IgGMulti-class antibodies規(guī)格： 0.2ml

GLUT2(Glucose transporter type 2) 葡萄糖轉(zhuǎn)運蛋白2(抗原)Multi-class antibodies規(guī)格： 0.5mg

孕激素誘導阻斷因子抗體 Anti-PIBF 0.2ml

14-3-3 sigma 英文名稱： 14-3-3σ抗體 0.2ml

EDNRA 英文名稱： 內(nèi)皮素受體A抗體 0.1ml

Rhesus antibody Rh phospho-MYL6(Tyr29+Ser30) 磷酸化肌球蛋白輕鏈6抗體 規(guī)格 0.1ml

GLUT2(Glucose transporter type 2) 葡萄糖轉(zhuǎn)運蛋白2(抗原)Multi-class antibodies規(guī)格： 0.5mg

Anti-PR/FITC 熒光素標記孕激素受體抗體IgGMulti-class antibodies規(guī)格： 0.2ml

Rabbit Anti-chicken IgM Whole serum 兔抗雞IgM抗血清Multi-class antibodies規(guī)格： 1ml

胰島素樣生長因子結合蛋白-2抗體 Anti-IGFBP-2 0.2ml

Rabbit Anti-chicken IgG 兔抗雞IgG 1mg

FYTTD1 英文名稱： FYTTD1蛋白抗體 0.2ml

Rhesus antibody Rh phospho-RAC1(Ser71) 磷酸化細胞遷移誘導因子5抗體 規(guī)格 0.1ml

Rabbit Anti-chicken IgM Whole serum 兔抗雞IgM抗血清Multi-class antibodies規(guī)格： 1ml

LDH(Lactate Dehydrogenase) 乳酸脫氫酶抗原Multi-class antibodies規(guī)格： 0.5mg

Anti-Hepatocyte 人肝細胞單克隆抗體Multi-class antibodies規(guī)格： 0.1ml

Rhesus antibody Rh Phospho-Cas (Tyr165) 磷酸化細胞凋亡敏感性基因 規(guī)格 0.1ml

馬血清封閉液 10ml Gibco原料,2%血清

V.cholera Ogawa 英文名稱： 01群小川型霍亂弧菌抗體 0.2ml

phospho-DNA PKcs (Thr2638) 英文名稱： 磷酸化DNA依賴性蛋白激酶抗體 0.1ml

Anti-Hepatocyte 人肝細胞單克隆抗體Multi-class antibodies規(guī)格： 0.1ml

IL36A Protein Human 重組人 IL1F6 / IL36A 蛋白

小鼠成纖維細胞完全培養(yǎng)基 100mL

DPP7 Others Mouse 小鼠 DPP7 / DPPII / DPP2 人細胞裂解液 (陽性對照)

BALB/3T3 clone A31小鼠胚胎成纖維細胞 BALB/3T3 clone A31 mouse embryo fibroblasts DMEM培養(yǎng)基+10%FBS

CL-0362HS 683(人腦細胞)5×106cells/瓶×2

NCI-H358(人非小細胞細胞) 5×106cells/瓶×2 SH-SY5Y(神經(jīng)母細胞瘤細胞)

CM-R041大鼠肝星形細胞完全培養(yǎng)基100mL

IFNGR1 Others Rat 大鼠 IFNGR / IFNGR1 人細胞裂解液 (陽性對照)

人脊柱間充質(zhì)干細胞HVMSC

JEG-3/VP16細胞，人絨癌細胞耐藥株 新生牛眼Tenon's囊成纖維細胞,NBTF細胞 人羊膜間充質(zhì)基質(zhì)細胞HAMSC

IM-9(人外周血B淋巴細胞) 5×106cells/瓶×2

95-D(人高轉(zhuǎn)移細胞) 5×106cells/瓶×2 CM-H099人真皮淋巴上皮細胞完全培養(yǎng)基100mL 人胚;CCC-HPF-1

AFG3樣蛋白2/脊髓小腦蛋白28抗體規(guī)格 IL36A Protein Human 重組人 IL1F6 / IL36A 蛋白

小鼠成纖維細胞完全培養(yǎng)基 100mL

DPP7 Others Mouse 小鼠 DPP7 / DPPII / DPP2 人細胞裂解液 (陽性對照)

BALB/3T3 clone A31小鼠胚胎成纖維細胞 BALB/3T3 clone A31 mouse embryo fibroblasts DMEM培養(yǎng)基+10%FBS

CL-0362HS 683(人腦細胞)5×106cells/瓶×2

NCI-H358(人非小細胞細胞) 5×106cells/瓶×2 SH-SY5Y(神經(jīng)母細胞瘤細胞)