抗体的生物素化标记实验要点：

1. AFG3样蛋白2/脊髓小脑蛋白28抗体规格 如在反应混合液中有叠氮钠或游离氨基存在,会抑制标记反应。因此,蛋白质在反应前要对 0.1mol/L碳酸氢钠缓冲液或0.5mol/L硼酸缓冲液充分透析；

2.所用的NHSB及待生物素化蛋白质之间的分子比按蛋白质表面的ε-氨基的密度会有所不同,选择不当则影响标记的效率,应先用几个不同的分子比来筛选最适条件；

3.用NHSB量过量也是不利的,抗原的结合位点可能因此被封闭,导致抗体失活；

4.由于抗体的氨基不易接近可能造成生物素化不足,此时可加入去污剂如 Triton x-100, Tween20等；

5.当游离ε-氨基(赖氨酸残基的氨基)存在于抗体的抗原结合位点时,或位于酶的催化位点时,生物素化会降低或损伤抗体蛋白的结合力或活性；

6.生物素还可能与不同的功能基团,如羰基、氨基、巯基、异咪唑基及苯酚基,也可与糖基共价结合；

7.交联反应后,应充分透析,否则,残余的生物素会对生物素化抗体与亲和素的结合产生竞争作用；

8.在细胞的荧光标记实验中,中和亲和素的本底低,但由于链霉亲和素含有少量正电荷,故对某些细胞可导致高本底。

产品订购信息：
英文名称  Anti-SCA28/AFG3L2 

中文名称   AFG3样蛋白2/脊髓小脑蛋白28抗体规格 

别    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 .

浓    度   1mg/1ml

规 格  0.2ml/200μg

抗体来源   Rabbit

克隆类型   polyclonal

交叉反应   Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Horse, Sheep 

产品类型   一抗  

研究领域     细胞生物 神经生物学 信号转导 泛素

蛋白分子量  predicted molecular weight: 89kDa

性    状   Lyophilized or Liquid

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

亚    型   IgG

纯化方法   affinity purified by Protein A

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

AFG3样蛋白2/脊髓小脑蛋白28抗体规格 产品应用    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

（石蜡切片需做抗原修复） 

 not yet tested in other applications.

 optimal dilutions/concentrations should be determined by the end user.  

保存条件  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. 

产品介绍 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

抗体的鉴定：

1）AFG3样蛋白2/脊髓小脑蛋白28抗体规格 抗体的效价鉴定：不管是用于诊断还是用于,制备抗体的目的都是要求较高效价。不同的抗原制备的抗体,要求的效价不一。鉴定效价的方法很多,包括有试管凝集反应,琼脂扩散试验,酶联免疫吸附试验等。常用的抗原所制备的抗体一般都有约成的鉴定效价的方法,以资比较。如制备抗抗体的效价,一般就采用琼脂扩散试验来鉴定。

2）抗体的特异性鉴定：抗体的特异性是指与相应抗原或近似抗原物质的识别能力。抗体的特异性高,它的识别能力就强。衡量特异性通常以交叉反应率来表示。交叉反应率可用竞争抑制试验测定。以不同浓度抗原和近似抗原分别做竞争抑制曲线,计算各自的结合率,求出各自在IC50时的浓度,并按公式计算交叉反应率。 

如果所用抗原浓度IC50浓度为pg/管,而一些近似抗原物质的IC50浓度几乎是无穷大时,表示这一抗血清与其他抗原物质的交叉反应率近似为0,即该血清的特异性较好。

3）抗体亲和力：是指抗体和抗原结合的牢固程度。亲和力的高低是由抗原分子的大小,抗体分子的结合位点与抗原决定簇之间立体构型的合适度决定的。有助于维持抗原抗体复合物稳定的分子间力有氢键,疏水键,侧链相反电荷基因的库仑力,范德华力和空间斥力。亲和力常以亲和常数K表示,K的单位是L/mol。抗体亲和力的测定对抗体的筛选,确定抗体的用途,验证抗体的均一性等均有重要意义。

Anti-STAT3 /FITC 荧光素标记信号转导和转录激活因子3抗体IgGMulti-class antibodies规格： 0.2ml

GLUT2(Glucose transporter type 2) 葡萄糖转运蛋白2(抗原)Multi-class antibodies规格： 0.5mg

孕激素诱导阻断因子抗体 Anti-PIBF 0.2ml

14-3-3 sigma 英文名称： 14-3-3σ抗体 0.2ml

EDNRA 英文名称： 内皮素受体A抗体 0.1ml

Rhesus antibody Rh phospho-MYL6(Tyr29+Ser30) 磷酸化肌球蛋白轻链6抗体 规格 0.1ml

GLUT2(Glucose transporter type 2) 葡萄糖转运蛋白2(抗原)Multi-class antibodies规格： 0.5mg

Anti-PR/FITC 荧光素标记孕激素受体抗体IgGMulti-class antibodies规格： 0.2ml

Rabbit Anti-chicken IgM Whole serum 兔抗鸡IgM抗血清Multi-class antibodies规格： 1ml

胰岛素样生长因子结合蛋白-2抗体 Anti-IGFBP-2 0.2ml

Rabbit Anti-chicken IgG 兔抗鸡IgG 1mg

FYTTD1 英文名称： FYTTD1蛋白抗体 0.2ml

Rhesus antibody Rh phospho-RAC1(Ser71) 磷酸化细胞迁移诱导因子5抗体 规格 0.1ml

Rabbit Anti-chicken IgM Whole serum 兔抗鸡IgM抗血清Multi-class antibodies规格： 1ml

LDH(Lactate Dehydrogenase) 乳酸脱氢酶抗原Multi-class antibodies规格： 0.5mg

Anti-Hepatocyte 人肝细胞单克隆抗体Multi-class antibodies规格： 0.1ml

Rhesus antibody Rh Phospho-Cas (Tyr165) 磷酸化细胞凋亡敏感性基因 规格 0.1ml

马血清封闭液 10ml Gibco原料,2%血清

V.cholera Ogawa 英文名称： 01群小川型霍乱弧菌抗体 0.2ml

phospho-DNA PKcs (Thr2638) 英文名称： 磷酸化DNA依赖性蛋白激酶抗体 0.1ml

Anti-Hepatocyte 人肝细胞单克隆抗体Multi-class antibodies规格： 0.1ml

IL36A Protein Human 重组人 IL1F6 / IL36A 蛋白

小鼠成纤维细胞完全培养基 100mL

DPP7 Others Mouse 小鼠 DPP7 / DPPII / DPP2 人细胞裂解液 (阳性对照)

BALB/3T3 clone A31小鼠胚胎成纤维细胞 BALB/3T3 clone A31 mouse embryo fibroblasts DMEM培养基+10%FBS

CL-0362HS 683(人脑细胞)5×106cells/瓶×2

NCI-H358(人非小细胞细胞) 5×106cells/瓶×2 SH-SY5Y(神经母细胞瘤细胞)

CM-R041大鼠肝星形细胞完全培养基100mL

IFNGR1 Others Rat 大鼠 IFNGR / IFNGR1 人细胞裂解液 (阳性对照)

人脊柱间充质干细胞HVMSC

JEG-3/VP16细胞，人绒癌细胞耐药株 新生牛眼Tenon's囊成纤维细胞,NBTF细胞 人羊膜间充质基质细胞HAMSC

IM-9(人外周血B淋巴细胞) 5×106cells/瓶×2

95-D(人高转移细胞) 5×106cells/瓶×2 CM-H099人真皮淋巴上皮细胞完全培养基100mL 人胚;CCC-HPF-1

AFG3样蛋白2/脊髓小脑蛋白28抗体规格 IL36A Protein Human 重组人 IL1F6 / IL36A 蛋白

小鼠成纤维细胞完全培养基 100mL

DPP7 Others Mouse 小鼠 DPP7 / DPPII / DPP2 人细胞裂解液 (阳性对照)

BALB/3T3 clone A31小鼠胚胎成纤维细胞 BALB/3T3 clone A31 mouse embryo fibroblasts DMEM培养基+10%FBS

CL-0362HS 683(人脑细胞)5×106cells/瓶×2

NCI-H358(人非小细胞细胞) 5×106cells/瓶×2 SH-SY5Y(神经母细胞瘤细胞)