关键词:抗体制备抗体标记重组蛋白表达

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多克隆抗体制备服务


服务介绍

多克隆抗体在免疫学诊断领域应用广泛。当机体受到抗原刺激后, 产生免疫学反应,由机体的浆细胞合成并分泌的与抗原有特异性结合能力的一组球蛋白(免疫球蛋白),这种与抗原有特异性结合能力的免疫球蛋白就是抗体。 抗体的特异性取决于抗原分子的决定簇,各抗原分子具有许多抗原决定簇。因此,由它免疫动物所产生的抗血清实际上是多种抗体的混合物,称为多克隆抗体。相对于单抗来说,它能识别多个抗原表位。由于具有可识别多个表位、可引起凝集反应、沉淀反应等优点,被广泛应用于免疫学诊断领域。

福因德科技(FrdBio)以提供技术服务为核心,专注从事抗体制备服务。企业围绕抗体生产与检测平台建立了多肽合成实验中心、重组蛋白定制实验中心和规范化动物养殖基地。根据客户所提供的起始材料,由经验丰富的抗原表位设计工程师设计并制定专项实验方案加以实施,最终提供给您符合您终实验需求的抗体。

 

Frdbio服务优势:

Ø以抗体最终应用为目的设计抗体制备路线,为客户提供一站式抗原-抗体制备方案。

Ø免疫采用自主研发的水溶性佐剂,大幅度提高抗体效价。

Ø效价保证:小分子/多肽类抗原ELISA效价≥1:105;蛋白类抗原ELISA效价≥1:106

 

服务要求(客户提交的材料)

蛋白、多肽、小分子化合物、菌体或基因序列信息等。

 

服务报告内容

1.抗原制备详细实验方案(若由基因序列起始);

2.抗原-抗体制备过程原始数据与抗体检测数据;

3.抗血清或纯化后的抗体。

 

服务编号、交付材料与周期

       本表中免疫宿主以新西兰大白兔为例,若客户要求宿主选择为波尔多羊,详情请咨询。

    定制抗体属于技术服务型产品,价格依据项目本身难易程度或当季优惠会有所浮动,具体以项目评估后项目设计方案双方商议为准。


套餐选项

服务内容

交付产品

周期

价格

多抗项目1

(从蛋白抗原到抗血清)

检测蛋白的纯度和浓度

SDS-PAGE胶检测图

8-10周

1880元

动物免疫及血清采集(2只)

1-2ml免疫前血清、60-80ml抗血清

ELISA检测

ELISA检测结果(针对免疫原)


完整项目报告

多抗项目2  

(从蛋白抗原到亲和纯化)

检测蛋白的纯度和浓度

SDS-PAGE胶检测图

10-12周

2600元

动物免疫及血清采集(2只)

1-2ml免疫前血清、纯化后剩余抗血清

亲和纯化

8-10mgA/G亲和纯化抗体

ELISA检测

ELISA检测结果(针对免疫原)


完整项目报告

多抗项目3  

(从多肽设计到抗血清)

多肽设计与合成(1条,12-22aa,90%)

2-3mg多肽、免疫后剩余多肽

12-15周

3580元

多肽偶联

多肽MS,HPLC图

动物免疫及血清采集(2只)

1-2ml免疫前血清、60-80ml抗血清

ELISA检测

ELISA检测结果(针对免疫原)


完整项目报告

多抗项目4

(从多肽设计到亲和纯化)

多肽设计与合成(1条,12-22aa,90%)

2-3mg多肽、免疫后剩余多肽

12-15周

4600元

多肽偶联

多肽MS,HPLC图

动物免疫及血清采集(2只)

1-2ml免疫前血清、纯化后剩余抗血清

亲和纯化

8-10mgA/G亲和纯化抗体

ELISA检测

ELISA检测结果(针对免疫原)


完整项目报告

多抗项目5

(磷酸化多抗制备)

多肽设计与合成(2条,12-22aa,95%)

2-3mg多肽(非磷酸化和磷酸化)

13-16周

评估

多肽偶联

多肽MS,HPLC图

抗原特异性亲和纯化柱制备

特异性纯化柱

动物免疫及血清采集(4只)

1-2ml免疫前血清

抗原特异性亲和纯化

3-5mg磷酸化抗体

ELISA检测

ELISA检测结果(针对免疫原)


完整项目报告

多抗项目6

(Western检测阳性多抗制备)

多肽设计与合成(2条,12-22aa,90%)或蛋白原核表达

2-3mg多肽或原核表达蛋白1mg

15-22周

评估

多肽偶联或基因调取,构建,原核表达蛋白

多肽MS,HPLC图或重组质粒,表达菌株

动物免疫及血清采集(4只)

1-2ml免疫前血清、纯化后剩余抗血清

抗原特异性亲和纯化

3-5mg抗原特异性亲和纯化抗体

ELISA检测

ELISA检测结果(针对免疫原)

Western Blot检测

Western Blot检测结果


完整项目报告

多抗项目7

(IHC检测阳性多抗制备)

多肽设计与合成(2条,12-22aa,90%)或蛋白原核表达

2-3mg多肽或原核表达蛋白1mg

15-22周

评估

多肽偶联或基因调取,构建,原核表达蛋白

多肽MS,HPLC图或重组质粒,表达菌株

动物免疫及血清采集(4只)

1-2ml免疫前血清、纯化后剩余抗血清

抗原特异性亲和纯化

3-5mg抗原特异性亲和纯化抗体

ELISA检测

ELISA检测结果(针对免疫原)

IHC检测

IHC检测结果(客户提供切片,修复条件)


完整项目报告

多抗项目8

(Western和IHC检测阳性多抗制备)

多肽设计与合成(4条,12-22aa,90%)或蛋白表达

2-3mg多肽或原核表达蛋白1mg

15-22周

评估

多肽偶联或基因调取,构建,原核表达蛋白

多肽MS,HPLC图或重组质粒,表达菌株

动物免疫及血清采集(8只)

1-2ml免疫前血清、纯化后剩余抗血清

抗原特异性亲和纯化

3-5mg抗原特异性亲和纯化抗体

ELISA检测

ELISA检测结果(针对免疫原)

Western Blot,IHC检测

Western Blot,IHC检测结果


完整项目报告

 

备注:

1.除以上常用的保证型多抗制备外,还可提供甲基化、乙酰化、羰基化、IP、Chip等其他类型的多抗制备,请咨询售前客服或销售人员;

2.多肽设计合成与重组蛋白表达详情请参照抗原制备;

多克隆抗体制备

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典型案例1:

Phosphatidylinositol synthesis, its selective salvage, and inter-regulation of anionic phospholipids in Toxoplasma gondii

Bingjian Ren Pengfei Kong Fatima Hedar Jos F Brouwers Nishith Gupta 

Commun Biol.2020 Dec 10;3(1):750. doi: 10.1038/s42003-020-01480-5.

Methods:

Biological reagents and resources. The RHΔku80Δhxgprt 75 and

RHΔku80Δhxgprt-TIR1 49 strains of T. gondii were kindly offered by Vern Carruthers (University of Michigan, MI) and David Sibley (Washington State University, St. Louis, MO), respectively. The Δtgcds1 r mutant was generated in our earlier study 7 . The pG140 plasmid was donated by Markus Meissner (Ludwig-Maximilians University, Munich). Antibodies recognizing the TgHSP90, TgGAP45,TgPDH-E1a, TgSAG2, and TgIMC3 proteins were bestowed by Sergio Angel (IIB-INTECH, Argentina), Dominique Soldati-Favre (University of Geneva, Switzer-land), Bang Shen (Huazhong Agricultural University, China), Honglin Jia (HarbinVeterinary Research Institute, China) and Marc-Jan Gubbels (Boston College,MA), respectively. Anti-TgSERCA and anti-TgPSD1mt antisera were obtained from FriendBio Bioscience and Technology (China). Other primary antibodies recognizing the TgSAG1 protein and engineered epitopes (HA and Ty1) were obtainedfrom ThermoFisher Scientific and Sigma-Aldrich (Germany). Corresponding secondary antibodies (Alexa488, Alexa594) were procured from Life Technologies(Germany). [ 3 H]-myo-inositol, and silica plates for TLC were obtained from American Radiolabeled Chemicals (St. Louis, MO) and Merck Millipore (Billerica,Fig. 9 Model of PtdIns biosynthesis and homeostasis of anionic phospholipids in T. gondii. PtdIns is made by TgPIS in the Golgi network using CDP-DAGand myo-inositol. CDP-DAG is generated by TgCDS1 in the endoplasmic reticulum, whereas myo-inositol is imported from the milieu. De novo synthesis ofmyo-inositol involves conversion of glucose to glucose-6-phosphate by hexokinase, followed by the action of inositol-3P synthase to produce inositol-3-phosphate. Successively, Ins3P is dephosphorylated to make inositol by inositol-3P monophosphatase. There is no biochemical and genetic evidence for the latter enzyme in T. gondii. Knockdown of TgPIS reduces the level of many PtdIns species along with homeostatic modulation of other anionic lipids (namelyPtdThr, PtdSer, PtdGro species). PtdIns species with shorter chains (e.g., C30/C32/C34) are synthesized de novo, while PtdIns species with relatively longer acyl chain (C38/C40) are salvaged by the parasite from its host cell. Based on isotope labeling studies, we speculate that both routes contribute to the synthesis of C36 PtdIns species (not shown). Transporters, inter-organelle contact sites, or other lipid trafficking pathways, as stated, enable transport of myo-inositol and CDP-DAG. The question-marked reaction specifies yet-unknown enzyme involved in CDP-DAG-dependent PtdSer synthesis. Selected abbreviations: CDS, CDP-DAG synthase; PGPS, PtdGro phosphate synthase; PGPP, PtdGro phosphate phosphatase; PSS, PtdSer synthase; PTS, PtdThrsynthase; PPM, parasite’s plasma membrane; PVM, parasitophorous vacuole membrane.

MA), respectively. [ 13 C]-myo-inositol was kindly provided by Dorothea Fiedler(Leibniz Research Institute, Berlin). Lipid standards and 1,2-dioleoyl-sn-glycero-3-(cytidine diphosphate) (CDP-DAG) were delivered by Avanti Polar Lipids (USA).DNA oligonucleotides (Supplementary Table 2) were purchased from Life Technologies.

 

典型案例2:

Functional evaluation of bacterial surface displayed P domain from human norovirus capsid proteins

S Rong, Y Zhou, M Niu, Q Li, M Wang… - … Material Science …, 2016 - atlantis-press.com

… Anti-VP1 (GI.1) or anti-VP1 (GII.4) polyclonal antisera (Friendbio Science and Technology (Wuhan) Co., Ltd., Hubei, China) were used as primary antibodies. The horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G (IgG) was used as secondary antibodies …

 

典型案例3:

Immunoblotting Quantification Approach for Identifying Potential Hypoallergenic Citrus Cultivars

J Wu, W Deng, D Lin, X Deng, Z Ma - Journal of agricultural and …, 2018 - ACS Publications

Production of Rabbit Polyclonal Antibodies. In order to generate antibodies against Cit s 1.01, Cit s 2.01, and Cit s 3.01, two New Zealand white rabbits (purchased from Hubei Provincial Center for Disease Control and Prevention, China) were immunized subcutaneously with 0.1 mg of recombinant GST-(Cit s 1.01)×2, GST-(Cit s 2.01)×3, and Trx-His-(Cit s 3.01)×2 proteins, respectively. Immunization and serum collection were performed by Friendbio Science & Technology (Wuhan) Co., Ltd. The protocol was approved by the Institute of Animal Care and Use Committee (no.201703083), according to national guidelines for animal care and handling, China. Immunization was performed every 2 weeks for a period of 3 months. The sera were collected after 3 months of immunization. Antibody titers were measured by ELISA. The sera were stored at 80 °C for further use


典型案例4:

Functional Divergence of Multiple Duplicated Foxl2 Homeologs and Alleles in A Recurrent Polyploid fish

RH Gan, Y Wang, Z Li, ZX Yu, XY Li… - Molecular Biology …, 2021 - academic.oup.com

Polyclonal antibody preparation, Western blot, histological section and immunofluorescence Polyclonal antibodies respectively specific to the CgFoxl2a and CgFoxl2b were produced by Friendbio Science & Technology (Wuhan) Co,.Ltd. Western blot,histological section and immunofluorescence were performed as described previously (Liu et al. 2015; Yang et al. 2017), and the results were acquired with ImageQuant LAS 4000mini (GE) and Carl Zeiss upright fluorescence microscope Axio Imager M2 (Analytical & Testing Center, IHB, CAS), respectively.


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tech@friendbio.com


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