Neuropeptide Y (NPY) is a member of a regulatory peptide family and has a marked sequence homology with pancreatic polypeptide (PP) and peptide YY (PYY), which are other members of the family. NPY is widely expressed in the nervous system and has been shown to be differentially expressed in inhibitory interneurons in the hippocampus in degenerative disease, as a powerful vasoconstrictor in the periphery, and increased expression of NPY in the hypothalamus correlates with food intake. In the rat central nervous system, immunohistochemistry has found NPY-like cell bodies in the cortex, caudate-putamen, hypothalamus (arcuate nucleus), hippocampus, anterior westolfactory bulb, nucleus accumbens, amygdaloid complex and periaqueductal grey. NPY-like fibers and terminals are detected in high numbers in the bed nucleus of the stria terminalis, the peri- and paraventricular regions of the hypothalamus and thalamus and in discrete hypothalamic nuclei, particularly the suprachiasmatic nucleus.
Westerns are not recommended for the Neuropeptide Y antibody since the protein is too small to detect in gel.
Photo Description: IHC image of neurons staining for neuropeptide Y in the rat cortex (above) and striatum (below). The tissue was fixed with 4% formaldehyde in phosphate buffer, before being removed and prepared for vibratome sectioning. Floating sections were incubated at RT in 10% goat serum in PBS, before standard IHC procedure. Primary antibody was incubated at 1:5000 for 48 hours, goat anti-rabbit secondary was subsequently added for 1 hour after washing with PBS. Light microscopy staining was achieved with standard biotin-streptavidin/HRP procedure and DAB chromogen. The second image above entitled "C." is NPY interneurons (RED) in mouse striatum, GREEN staining is GFP expressed by medium spiny neurons provided Alipi Naydenov, Neurobiology and Behavior, Stella Lab University of Washington (see review below).
Host: Rabbit
Quantity / Volume: 100 µL
State: Lyophilized Whole Serum
Reacts With: Alpaca (Llama), Bird, Bubalus Bubalis (Water Buffalo), Bullfrog, Cat, Ewe, Ferret, Fish, Frog, Frog (Xenopus Laevis), Ground Squirrel, Guinea Pig, Hamster, Human, Monkey, Mouse, Ovine (Sheep), Pig, Psetta Maxima (Flat Fish), Rat, Sheep, Snake, Tadpole, Toad, Turtle, Zebrafish
Availability: In Stock
Preabsorption Control: Available at Sigma Aldrich, Catalog # N3266
Alternate Names: Pro-neuropeptide Y; Neuropeptide tyrosine; prepro-neuropeptide Y; PYY4, anti-NPY
Gene Symbol: NPYRRID: AB_572253
Database Links:
Entrez Gene: 102414548 Bubalus Bubalis (Water Buffalo) Entrez Gene: 101096968 Cat Entrez Gene: 443508 Ewe Entrez Gene: 101686990 Ferret Entrez Gene: 779985 Frog Entrez Gene: 101976112 Ground Squirrel Entrez Gene: 100379621 Guinea Pig Entrez Gene: 100758436 Hamster Entrez Gene: 4852 Human Entrez Gene: 574114 Monkey Entrez Gene: 109648 Mouse Entrez Gene: 397304 Pig Entrez Gene: 24604 Rat Entrez Gene: 102454441 Turtle Entrez Gene: 30281 Zebrafish
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1.单克隆抗体的优点:
(1)杂交瘤可以在体外“永久”地存活并传代,只要不发生细胞株的基因突变,就可以不断的生产高特异性、高均一性的抗体.
(2)可以用相对不纯的抗原,获得大量高度特异的、均一的抗体.
(3)由于可能得到“无限量”的均一性抗体,所以适用于以标记抗体为特点的免疫学分析方法,如IRMA和ELISA等.
(4)由于单克隆抗体的高特异性和单一生物学功能,可用于体内的放射免疫显像和免疫导向治疗.
2.单克隆抗体的局限性:
(1)单克隆抗体固定的亲和性和局限的生物活性限制了它的应用范围.由于单克隆抗体不能进行沉淀和凝集反应,所以很多检测方法不能用单克隆抗体完成.
(2)单克隆抗体的反应强度不如多克隆抗体.
(3)制备技术复杂,而且费时费工,所以单克隆抗体的价格也较高.
抗原有两个基本特性,即抗原性和免疫原性。有抗原性的物质不一定有免疫原性,所以由此引出半抗原和完全全抗原,半抗原必须经过经过一定的改造(偶联蛋白载体BSA,OVA或者HSA等大分子物质)方能成为完全。一般而言完全抗原分子量越大(大于10KDa),结构越复杂引起免疫反应的能力也就越强。
抗体就是能与特异性抗原结合的免疫球蛋白,抗体一般分为多克隆抗体和单克隆抗体,多克隆抗体能与抗原的多个表位结合。本篇主要讲述兔来源的多克隆抗体的生产步骤
多抗一般制备流程:完全抗原的准备→兔子的免疫→ 效价检测和终放→抗体亲和纯化→抗体的浓缩和保存。
又由于自然存在的抗原大都存在多个抗原表位,会刺激机体产生多种针对同一抗原的不同抗原表位相应的不同抗体.
一般来说多克隆的阳性率高一些,但出现假阳性的比例也高一些。
其次,察看次目的蛋白的存在形式,有没有多聚体形式及变构形式;
最后,查看多家抗体公司的DATA,看看别人的WB做出来的条带的位置。
根据你说的,特异识别多个组织中的同样大小的条带,我觉得很可能就是你的目的蛋白。
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