抗体来源 Rabbit
克隆类型 polyclonal
交叉反应 Human, Mouse, Rat, Chicken, Dog, Pig, Cow, Sheep
产品类型 一抗
研究领域 细胞生物 神经生物学 信号转导 转录调节因子 G蛋白偶联受体 表观遗传学 G蛋白信号
蛋白分子量 predicted molecular weight: 209kDa
性 状 Lyophilized or Liquid
免 疫 原 KLH conjugated synthetic peptide derived from human BIG1/ARFGEF1
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
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.
ARF鸟苷酸交换因子BIG1抗体产品介绍 Guanine nucleotide-exchange proteins (GEPs) accelerate replacement of bound GDP with GTP and thereby activate ADP-ribosylation factors (ARFs), a family of guanine nucleotide-binding proteins that play an important role in intracellular vesicular trafficking. GEPs comprise two major families, large GEPs that are inhibited by brefeldin A (BFA), a protein that effects Golgi structure and a group of smaller GEPs that are insenstive to BFA. Two genes for GEPs found on human chromosomes 8 and 20 encode BFA sensitive GEPs designated BIG1 and BIG2. Both GEPS contain a sec7 domain that is responsible for their brefeldin inhibition and also their catalytic activity. In vivo, BIG1 and BIG2 exist in macromolecular complexes that move between the Golgi membranes and cytosol. BIG2 associates with PKA regulatory subunits, implying that BIG2 may act as an A kinase-anchoring protein (AKAP) that could coordinate the cAMP and ARF regulatory pathways.
Function : BIG1 is a brefeldin A-inhibited guanine nucleotide exchange protein (GEP) that activates ADP-ribosylation factor (ARF) GTPases and functions in vesicle formation from Golgi membranes.
Subunit : Homodimer (Probable). Interacts with BIG1; both proteins are probably part of the same or very similar macromolecular complexes. Interacts with FKBP2, DPY30, MYO9B, PRKAR1A, PRKAR2A, PPP1CC and PDE3A. Interacts with KANK1; however, colocalization cannot be experimentally confirmed. Interacts with NCL, FBL, NUP62 and U3 small nucleolar RNA.
Subcellular Location : Cytoplasm. Cytoplasm, perinuclear region. Golgi apparatus. Golgi apparatus, trans-Golgi network (By similarity). Nucleus. Nucleus, nucleolus. Nucleus matrix. Membrane. Note=Translocates from cytoplasm to membranes and nucleus upon cAMP treatment.
Tissue Specificity : Expressed in placenta, lung, heart, brain, kidney and pancreas.
Post-translational modifications : Phosphorylated. In vitro phosphorylated by PKA reducing its GEF activity and dephosphorylated by phosphatase PP1.
Similarity : Contains 1 SEC7 domain
Database links : UniProtKB/Swiss-Prot: Q9Y6D6.2
纯度:在实验的任何阶段,确定抗体溶液纯度的最简单方法是取一部分样本进行SDS-PAGE电泳。凝胶可用考马斯亮蓝染色(灵敏度为0.1—0.5ug/带)或银染(灵敏度1~l0ug/带)。
定量:如果抗体还不纯,有一个快捷的定量方法,即通过SDS-PAGE电泳分离出轻、重链,然后和已知的标准染色带比较。如果需要分析许多样本,用免疫测定法对抗体定量较容易。如果抗体是经过纯化的,可通过测蛋白总量代替上述两种方法,有一简单的方法,即紫外吸收法。ARF鸟苷酸交换因子BIG1抗体的量可通过测280nm处的吸收值来测(10D大致相当于0.75mg/m1的纯化抗体)。
抗原结合活性:一般说来,纯化方法不会引起抗原结合活性的改变。用蛋白G或蛋白A树脂很少导致抗体活性丧失。然而,如果最终抗体产物的作用不如原来所预料的好,检测抗体纯化过程所丢失的活性就极为重要。用一系列滴定法比较纯化的抗体和其原材料的活性,以标定每一步中的总抗体量,这将有助于较好的估计通过纯化所丢失的活性。