![NSJ Bio/ABO Antibody / Blood Group Antigen B [clone HEB-29] (V2551)/100 ug/V2551SAF-100UG](images/no_picture.gif)
The antibody HEB-29 reacts with human blood group B. The specificity of the antibody HEB-29 was confirmed by comparison of specificity and reactivity to standard reagent using >5.000 samples of blood. mAb HEB-29 shows specific staining of erythrocytes and vascular epithelium of blood group B controls and no staining in group A controls. It is applicable for tissue staining in tumor patients with blood groups B and AB. Blood group antigens are generally defined as molecules formed by sequential addition of saccharides to the carbohydrate side chains of lipids and proteins detected on erythrocytes and certain epithelial cells. The A, B and H antigens are reported to undergo modulation during malignant cellular transformation. Blood group related antigens represent a group of carbohydrate determinants carried on both glycolipids and glycoproteins. They are usually mucin type, and are detected on erythrocytes, certain epithelial cells, and in secretions of certain individuals. Sixteen genetically and biosynthetically distinct but inter related specificities belong to this group of antigens, including A, B, H, Lewis A, Lewis B, Lewis X, Lewis Y, and precursor type 1 chain antigens.
Optimal dilution of the ABO antibody should be determined by the researcher.
1. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM Citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes
A mixture of erythrocytes of group B and glycoprotein fraction isolated from saliva of secretors with blood group B was used as the immunogen for the ABO antibody.
Store the ABO antibody at 2-8oC (with azide) or aliquot and store at -20oC or colder (without azide).
NSJBio为生命科学界提供经典,广泛使用的单克隆抗体和的,新推向市场的单克隆和多克隆抗体,以及重组单克隆和人类蛋白质微阵列验证抗体的来源研究领域包括癌症,肿瘤生物标志物,细胞凋亡,免疫学,细胞生物学,转录因子和表观遗传学。许多产品提供多种尺寸,以更好地适应研究人员的测试需求和预算限制。全面的产品数据表提供产品信息和测试数据图像,并由经验丰富的技术服务提供支持。并且所有产品100%保证按照产品数据表中的说明进行操作。
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NSJBio | R35717-100UG | DCN1Antibody | 100ug |
NSJBio | R35718-100UG | DAOAntibody | 100ug |
NSJBio | R35719-100UG | ZEB1Antibody | 100ug |
NSJBio | R35720-100UG | NPFFR1Antibody | 100ug |
NSJBio | R35721-100UG | TZFPAntibody | 100ug |
NSJBio | R35724-100UG | MFSD6Antibody | 100ug |
NSJBio | R35728-100UG | ARPC3Antibody | 100ug |
NSJBio | R35729-100UG | P15INK4bAntibody(isoform2) | 100ug |
NSJBio | R35730-100UG | DKK4Antibody | 100ug |
NSJBio | R35731-100UG | PITPNBAntibody | 100ug |
NSJBio | R35732-100UG | MUNC18CAntibody | 100ug |
NSJBio | R35736-100UG | PSPHAntibody | 100ug |
NSJBio | R35737-100UG | CD123Antibody | 100ug |
NSJBio | R35738-100UG | Wfdc1Antibody | 100ug |
NSJBio | R35739-100UG | RXRGAntibody | 100ug |
NSJBio | R35741-100UG | TRIM71Antibody | 100ug |
NSJBio | R35742-100UG | FAFAntibody | 100ug |
NSJBio | R35744-100UG | Cntn4Antibody | 100ug |
NSJBio | R35747-100UG | EPB41L2Antibody | 100ug |
NSJBio | R35748-100UG | P16INK4aAntibody | 100ug |
NSJBio | R35749-100UG | AIDAntibody | 100ug |
NSJBio | R35750-100UG | RIG1Antibody | 100ug |
NSJBio | R35751-100UG | OXTRAntibody | 100ug |
NSJBio | R35752-100UG | IL-21Antibody | 100ug |
NSJBio | R35754-100UG | ImportinbetaAntibody | 100ug |
NSJBio | R35755-100UG | PRKCDBPAntibody | 100ug |
NSJBio | R35756-100UG | GPR83Antibody | 100ug |
NSJBio | R35757-100UG | HES4Antibody | 100ug |
NSJBio | R35758-100UG | CRLS1Antibody | 100ug |
NSJBio | R35759-100UG | CrotAntibody | 100ug |
NSJBio | R35760-100UG | UNC45BAntibody | 100ug |
NSJBio | R35762-100UG | LAT2Antibody | 100ug |
NSJBio | R35763-100UG | POU6F2Antibody | 100ug |
NSJBio | R35764-100UG | GLAST1Antibody | 100ug |
NSJBio | R35765-100UG | ProenkephalinAntibody | 100ug |
NSJBio | R35767-100UG | FibrinogenbetachainAntibody | 100ug |
NSJBio | R35768-100UG | GALR1Antibody | 100ug |
NSJBio | R35770-100UG | TMX1Antibody | 100ug |
NSJBio | R35770BTN | TMX1Antibody[BiotinConjugate] | 100ug |
NSJBio | R35772-100UG | ACADVLAntibody | 100ug |
NSJBio | R35773-100UG | VHLAntibody | 100ug |
NSJBio | R35775-100UG | HCSTAntibody | 100ug |
NSJBio | R35776-100UG | CebpzAntibody | 100ug |
NSJBio | R35778-100UG | CHCHD3Antibody | 100ug |
NSJBio | R35779-100UG | TTLL4Antibody | 100ug |
NSJBio | R35780-100UG | TCF19Antibody | 100ug |
NSJBio | R35781-100UG | IGF1Antibody | 100ug |
NSJBio | R35782-100UG | WIZAntibody | 100ug |
NSJBio | R35783-100UG | GOT1Antibody | 100ug |
NSJBio | R35784-100UG | FMR1Antibody | 100ug |
NSJBio | R35785-100UG | UPARAntibody | 100ug |
NSJBio | R35786-100UG | Peptidaseinhibitor15Antibody | 100ug |
NSJBio | R35787-100UG | FURINAntibody | 100ug |
NSJBio | R35788-100UG | AxotrophinAntibody | 100ug |
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NSJBio | R35796-100UG | SPON2Antibody | 100ug |
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NSJBio | R35799-100UG | KIAA1881Antibody | 100ug |
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鼠源性抗体应用于人体会产生抗鼠抗体反应(HAMA )等,妨碍其在临床上的应用,所以要制备人鼠杂交和完全人源化的抗体,减少抗体中的鼠源成分,尽量保留原有抗体的特异性.
希望得到前辈们的回答。非常感谢!
单克隆抗体是利用了瘤细胞的无限增殖,利用了淋巴细胞能产生单一抗体的特征。记忆细胞是不能产生抗体的,需要抗原刺激才能增殖、分化为浆细胞,浆细胞才能产生抗体。
单克隆抗体这项新技术从根本上解决了在抗体制备中长期存在的特异性和可重复性问题,可用于探讨: ①蛋白质的精细结构;②淋巴细胞亚群的表面新抗原;③组织相容性抗原;④激素和药物的放射免疫(或酶免疫)分析;⑤肿瘤的定位和分类;⑥纯化微生物和寄生虫抗原;⑦免疫治疗和与药物结合的免疫-化学疗法 (“导弹”疗法,利用单克隆抗体与靶细胞特异性结合,将药物带至病灶部位.。
因此,单克隆抗体可直接用于人类疾病的诊断、预防、治疗以及免疫机制的研究,为人类恶性肿瘤的免疫诊断与免疫治疗开辟了广阔前景。
最近我想筛选一个针对膜蛋白的单克隆抗体,这个抗体的参考文献中提到的一种筛选方法是用DNA质粒直接免疫小鼠,然后再脾融合筛选出来的。这里有几个问题想向大家请教一下:
1.DNA效果比蛋白差很远,但是膜蛋白的抗体有可能是针对空间构象的,能起效的抗体位点很少,所以要在免疫阶段保证抗体多样性和特异性。看参考文献的剂量都是25ug或者50ug这种比较低的剂量,免疫次数2-3次,最后在融合前用细胞加免一次。而我设计的实验组是100ug剂量,免疫了4-5次,我这样是不是筛不出来特异性的抗体了?
2.DNA筛到的阳性抗体,可能亲和力低,如果不想筛到IgM的抗体,DNA免疫从什么时候开始由IgM转换成IgG的?抗体多样性在免疫的什么阶段比较高?
向左转|向右转
首先是一个抗体药物的releaselotstest:
?ProteinConcentration----------------------70.9mg/mL[63.0-77.0]
?Potency----------------------100%[80-125%]
?ID(immunoassay)----------------------Pass
?SE-HLC-----------------------MP:99.8%,HMW:0.2%,LMW:0.0%[MP≥99%]
?CE-SDS,reduced----------------------HC+LC:98.9%,HC:66.8%LC:32.1%[LC+HC≥98%]
?CEX-HPLC-----------------------MP:89.6%,Acidic:2.7%,Basic:7.7%[≥80%]
?CEX-HPLCID-----------------------Pass
?Osmolality-----------------------314mOsm/kg[300±50]
?pH----------------------5.2at24.6°C[5.0–5.4]
?Appearance-----------------------Report
?Volume-----------------------1.2mL(average1.1525mL,n=5)[≥1mL]
?Sub-visIBLeParticulate----------------------29particles/container≥10μm[nomorethan6000]
3particles/container≥25μm[nomorethan600]
?Sterility--------------------------------------Pass
?BacterialEndotoxin-----------------------<0.2EU/mL[≤5.0]
?CHOPELISA-----------------------4.4ppm
?Protein-AELISA-----------------------<0.5ppm
?DNAQPCR-----------------------<1pg/mg
以上各项指标基本是反映了对抗体药物最终产品的质量要求,用到的一些方法也比较的明了,可以作为抗体药物开发的参考。
版主zhulikou431留言:
加分鼓励有价值的话题。
实验外包单克隆抗体在现代生物医学中的作用越来越大,它不仅被广泛应用于生化和免疫检测,而且治疗癌症和自体免疫疾病的单抗药物也逐渐批准上市。因此单抗的制备和筛选具有其内在的重要性。
动物免疫
取4只6周龄雌性BALB/c小鼠皮下注射抗原(20μg/只),第一次免疫与等量弗氏完全佐剂乳化,0.2mL/只;两周后进行第二次免疫,抗原与等量弗氏不完全佐剂乳化,皮下注射,0.2mL/只;两周后进行第三次免疫,免疫方法及剂量同第二次免疫。第三次免疫一周后,小鼠眼眶采血测其效价,第三次免疫两周后,选择效价最高的小鼠,用不加佐剂的抗原加强免疫,3d后取小鼠脾细胞与SP2/0细胞融合。
细胞融合
1骨髓瘤细胞的准备
选择生长状态良好的SP2/0细胞,覆盖瓶底80%时弃上清,以不完全培养基洗涤一次后,用l0mL不完全培养基将细胞轻轻吹下。
2脾淋巴细胞的准备
取加强免疫后3天的小鼠,摘除眼球采血作为阳性血清;颈脱臼将小鼠致死,用75%酒精消毒体表10min,随即放入超净台内的解剖板上,腹部朝上,四肢固定;无菌打开腹腔取出脾脏,用基础培养基洗涤,并仔细去掉周围附着的结缔组织;随后将脾脏转移到另一个盛有DMEM的平皿中。用研磨棒挤压,使脾细胞充分释放,制成脾细胞悬液。
3饲养细胞的制备
取一只成熟健康的ICR小鼠,摘眼球采血作为阴性血清,颈脱臼处死;体表消毒和固定后,暴露腹膜,酒精棉球消毒腹膜;用l0mL注射器,12#针头,吸取10mLHAT培养基到腹腔,右手固定注射器,左手持酒精棉球轻轻按摩腹部,抽回腹腔内液体,注入己准备好的容器中。
4融合
将上述制备的骨髓瘤细胞与脾细胞混合于一支50mL的带盖的融合管中,1000rpm离心10min,弃上清。将融合管置于手掌中,轻轻摩擦底部,使两种细胞充分混匀;在37°C水浴中45s内缓慢加入1mL预热的PEG-1500到融合管中,边加边轻轻摇匀;随即在90s内先慢后快滴加30mL37°C预热的不完全DMEM培养基,使PEG稀释而失去作用,37°C静置10min,1000rpm离心10min;弃上清,用60mLHAT培养基轻轻悬浮沉淀细胞,再加入适量的腹腔巨噬细胞;分装于96孔细胞培养板,然后将培养板置37°C6%CO2培养箱内培养;5d后用新鲜的HAT培养基换出一半培养基;10d后用预热的HT换出HAT;观察杂交瘤细胞的生长情况,待其细胞培养上清变黄或克隆分布至孔底面积的1/10以上时,吸取适量细胞上清进行ELISA检测,两次检测结果为阳性设为阳性克隆孔。
杂交瘤细胞的克隆化及建株
采用有限稀释法对连续两次检测为阳性的细胞株进行克隆化。阳性细胞计数,取100个细胞放入5mL含饲养细胞的培养液中,即20个细胞/mL,100μL/孔滴加于96孔细胞培养板,即每孔1个细胞:再将余下的细胞悬液倍比稀释,细胞数为10个/mL,100μL/孔滴加于96孔细胞培养板,即每孔0.5个细胞。第8~9d时,肉眼可见细胞克隆,对出现单个细胞克隆的孔再按筛选阳性杂交瘤细胞的间接ELISA方法进行筛选。连续进行三次以上亚克隆,判为阳性的细胞株扩大培养,建株冻存。
体组织标本抗选种兔抗鼠抗类抗抗体
二抗针抗源抗兔抗二抗要选鼠抗兔或者羊抗兔类抗兔抗体(注意句抗兔抗基础抗鼠抗二抗要选抗鼠抗体)
我先开个头
泊落沙姆188,土温80

