Fab fragment antibodies are generated by papain digestion of whole IgG antibodies to remove the entire Fc portion, including the hinge region. These antibodies are monovalent, containing only a single antigen binding site. The molecular weight of Fab fragments is about 50 kDa.
Usage Physical State: Freeze-dried solidStorage and Rehydration: Store freeze-dried solid at 2-8°C. Rehydrate with the indicated volume of dH2O (see product specification sheet) and centrifuge if not clear. Prepare working dilution on day of use. Product is stable for about 6 weeks at 2-8°C as an undiluted liquid.Extended Storage after Rehydration: Aliquot and freeze at -70°C or below. Avoid repeated freezing and thawing. Alternatively, add an equal volume of glycerol (ACS grade or better) for a final concentration of 50%, and store at -20°C as a liquid. Expiration date: one year from date of rehydration. The expiration date may be extended if test results are acceptable for the intended use.
Purity:The antibody was purified from antisera by a combination of papain digestion and immunoaffinity chromatography using antigens coupled to agarose beads.Fc fragments and whole IgG molecules have been removed. Buffer: 0.01M Sodium Phosphate, 0.25M NaCl, pH 7.6 Stabilizer:15 mg/ml Bovine Serum Albumin (IgG-Free, Protease-Free) Preservative:0.05% Sodium Azide Suggested Working Concentration or Dilution Range:1:50 - 1:200 for sequential labeling applications.
To complex with primary antibody in solution, use 1:1 weight ratio of Fab:primary antibody (15:1 molar ratio). Vortex and incubate for 30 minutes at room temperature prior to use. Titrate complex to optimal dilution for assay.
Dilution factors are presented in the form of a range because the optimal dilution is a function of many factors, such as antigen density, permeability, etc. The actual dilution used must be determined empirically.
ConjugateRhodamine Red™-X (RRX)
Amax: 570Emax: 590nmRRX (Rhodamine Red-X) conjugates have a peak of excitation at 570 nm and a peak of emission at 590 nm. Although TRITC has been used traditionally with FITC for double labeling, better color separation is achieved by using RRX or Alexa Fluor® 594. Rhodamine Red-X is particularly useful for 3- and 4-color labeling with DyLight 405, Alexa Fluor® 488, and Alexa Fluor® 647 by using a confocal microscope equipped with a 405 nm laser and a krypton/argon laser. Fluorescence from RRX lies about midway between that of Alexa Fluor® 488 and Alexa Fluor® 647, and it shows little overlap with either dye. The krypton-argon laser emits lines at 488 nm, 568 nm, and 647 nm, which are optimal for exciting Alexa Fluor® 488, RRX, and Alexa Fluor® 647, respectively. By adding a 405 nm laser and a 420 nm emission filter, 4-color labeling is possible using DyLight 405-conjugated secondary antibodies from JIR (Figure 5). The separation between all four dyes is perfect for 4-color labeling, and all four dyes are very bright.
Images & ReferencesThis product is for in vitro research use only. It is not a medical device and it is not intended for diagnostic or therapeutic purposes.
Rhodamine Red-X is a trademark of Molecular Probes, Inc. Jackson ImmunoResearch is licensed by Molecular Probes, Inc. to manufacture and sell conjugates of Rhodamine Red-X reactive dye.
Jackson总部位于美国宾夕法尼的兰卡斯特,距离费城大概有40英里。Jackson在1982年由科学家展开了包括在细胞生物学,蛋白质化学,微生物学,免疫学及产品开发实验研究。我们的目标是为世界各地的研究者能最大选择的提供最高质量的科学试剂以及最棒的技术支持和售后服务。蚂蚁淘向您提醒的是,Jackson产品仅用于研究,不用于诊断或治疗用途。作为美国著名的各种二抗生产公司之一,Jackson在全世界各国设有几十家代理。由于其产品效价高、种类全、价格低廉,因而成为全球各大药厂、试剂盒生产厂、大型实验室采购此类试剂的首选。其产品主要包括:各种纯化的二抗、标记的二抗(AMCA、Cy2、FITC、Cy3、TRITC、RRX、TR、Cy5、长臂生物素、辣根酶和碱磷酶标记)、金标试剂等。
Jackson二抗产品选购简介
为便于客户选购Jackson公司的各种二抗,特将选购时的注意事项摘译如下:
JacksonImmunoResearch的抗体产品线完全,有全球最大的二抗产品库,能提供多种应用类型和需要的二抗产品,其二抗产品本身也具有极高的产品质量和稳定性。以上述产品为例,其HRP酶标山羊抗小鼠二抗,WB实验的应用稀释比率为1:10000-1:200000,也就是说,购买的2ml二抗,实际应用时相当于20L-400L的最终使用液。另外,一次购买大包装,避免了不同批次之前的差异,降低了实验条件本身的误差。作为多家著名品牌的代理商,蚂蚁淘根据国内科研的现实需求,为客户选择最具性价比的二抗及血清产品免疫学解决方案。如果您对Jackson的产品感兴趣,请致电027-84454412到蚂蚁淘科技有限公司问询您所感兴趣的产品类型。
JacksonImmunoResearchLaboratories,INC.专业致力于亲和纯化的二抗和免疫球蛋白的生产及偶联。产品主要销往世界各地的大学和科研院所。该公司位于宾夕法尼亚州费城以西约40英里,始建于1982年,经过40年左右的发展,Jackson的二抗以种类丰富(物种齐全,标记多样,应用广泛)、性价比高、供货迅捷而受到更多科研学者和工业客户的青睐。
Jackson的产品主要有酶标二抗、荧光标记二抗、生物素标记二抗、胶体金标记二抗、封闭血清等等,其中JacksonHRP标记羊抗兔/羊抗鼠二抗,更是很多生产厂家的重要物料之一。苏州蚂蚁淘生物科技有限公司作为Jackson在中国的代理商,备有大量二抗现货,并能提供专业的售前咨询和完善的售后服务。
JacksonImmunoResearchLaboratories,INC.专业致力于亲和纯化的二抗和免疫球蛋白的生产及偶联。产品主要销往世界各地的大学和科研院所。该公司位于宾夕法尼亚州费城以西约40英里,始建于1982年,经过40年左右的发展,Jackson二抗以种类丰富(物种齐全,标记多样,应用广泛)、性价比高、供货迅捷而受到更多科研学者和工业客户的青睐。
Jackson的所有产品均具有极高的纯度和高超的品质,大部分二抗产品以冻干粉形式存在,易于保存和运输;作为二抗的实际生产商(非OEM商),产品的性价比也是非常高的,很多大厂家诸如T.F等也会从JacksonOEM产品。另外厂家完善的库存和供应体系以及高效稳定的纯化技术,保证了大部分二抗产品均能现货形式供应,不同批次的产品质量的稳定性也非常高。
Jackson的产品主要有酶标二抗、荧光标记二抗、生物素标记二抗、胶体金标记二抗、封闭血清等等,其中JacksonHRP标记羊抗兔/羊抗鼠二抗,更是很多生产厂家的重要物料之一。苏州蚂蚁淘生物科技有限公司作为Jackson在中国代理商,备有大量二抗现货,并能提供专业的售前咨询和完善的售后服务。
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目前大量制备单抗的方法主要有两大系统,一是动物体内生产法,这是国内外实验室所广泛采用;另一是体外培养法。
(1)动物体内生产单抗的方法
迄今为止,通常情况下均采用动物体内生产单抗的方法,鉴于绝大多数动物用杂交瘤均由BALB/c小鼠的骨髓瘤细胞与同品系的脾细胞融合而得,因此使用的动物当然首选BALB/c小鼠。本方法即将杂交瘤细胞接种于小鼠腹腔内,在小鼠腹腔内生长杂交瘤,并产生腹水,因而可得到大量的腹水单抗且抗体浓度很高。可见该法操作简便、经济,不过,腹水中常混有小鼠的各种杂蛋白(包括Ig),因此在很多情况下要提纯后才能使用,而且还有污染动物病毒的危险,故而最好用SPF级小鼠。
(2)体外培养生产单抗的方法
总体上讲,杂交瘤细胞系并不是严格的贴壁依赖细胞(anchoragedependentcell,ADC),因此既可以进行单层细胞培养,又可以进行悬浮培养。杂交瘤细胞的单层细胞培养法是各个实验室最常用的手段,即将杂交瘤细胞加入培养瓶中,以含10-15%小牛血清的培养基培养,细胞浓度以1×106-2×106/ml为佳,然后收集培养上清,其中单抗含量约10-50ug/ml。显然,这种方法制备的单抗量极为有限,无疑是不适用于单抗的大规模生产。要想在体外大量制备单抗,就必须进行杂交瘤细胞的大量(高密度)培养。单位体积内细胞数量越多,细胞存活时间越长,单抗的浓度就越高,产量就越大。