- SourceDinitrophenyl (DNP) is a hapten that is not normally expressed in the target tissue. Therefore, anti-DNP antibody has no relevant specificity to a target antigen, and can be used as isotype control antibody to differentiate non-specific background signal from specific antibody signal in various in vitro and in vivo studies. Biotinylated Monoclonal Anti-DNP antibody, Mouse IgG1 Isotype Control was purified from HEK293 cell culture. It is the Biotin labeled form ofMonoclonal Anti-DNP antibody, Mouse IgG1 Isotype Control (Cat. No. DNP-M1).
- IsotypeMouse IgG1/kappa
- SpecificitySpecifically reacts with DNP (Dinitrophenyl) and DNP conjugated proteins.
- ApplicationThis antibody is suitable for use as a non-targeting isotype control in various in vitro and in vivo studies. It can also be used as a negative control in various applications such as ELISA, Western blot, immunofluorescence, immunohistochemistry, immunoprecipitation, and flow cytometry. Each laboratory should determine an optimum working titer for use in its particular application.
- Purity
>90% as determined by SDS-PAGE.
- Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 . Normally trehalose is added as protectant before lyophilization.
Contact us for customized product form or formulation.
- Reconstitution
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
- Storage
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.

Biotinylated Monoclonal Anti-DNP antibody, Mouse IgG1 Isotype Control on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

2e5 of Anti-CD19 CAR-293 cells were stained with a series of concentrations of Biotinylated Anti-DNP Antibody, Mouse IgG1 (Cat. No. DNP-BM1), washed and then followed by PE-SA and analyzed with FACS (QC tested).

5e5 of PBMCs were stained with a series of concentrations of Biotinylated Anti-DNP Antibody, Mouse IgG1 (Cat. No. DNP-BM1), washed and then followed by PE-SA and analyzed with FACS (QC tested).

2e5 of Anti-CD19 CAR-293 cells were stained with 100 μL of 1 μg/mL of Biotinylated Anti-DNP Antibody, Mouse IgG1 (Cat. No. DNP-BM1) and positive control respectively, washed and then followed by PE-SA and analyzed with FACS (Routinely tested).
- BackgroundA hapten is a small molecule that can elicit an immune response only when conjugated with a large carrier such as a protein. Typical haptens include drugs, urushiol, quinone, steroids, etc. Peptides and non-protein antigens usually need conjugating to a carrier protein (such as BSA (bovine serum albumin) or KLH (keyhole limpet hemocyanin) to become good immunogens). Additionally, haptens should be administered with an adjuvant to ensure a high quality immune response. It is important that the hapten design (preserving greatly the chemical structure and spatial conformation of target compound), selection of the appropriate carrier protein and the conjugation method are key conditions for the desired specificity anti-hapten antibodies. We design anti-hapten antibodies based on the HaptenDB information.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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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。
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。
1取组织
(1) 灌注固定:暴露心脏,将注射针头插入左心室,剪开右心耳,快速灌注PBS,待右心耳处流出澄清液体,换4%多聚甲醛灌注到大鼠四肢强直为止,快速取脑和肾。
(2) 固定:将组织置于4%多聚甲醛中固定15小时
(3) 脱水:将组织放入30% 蔗糖溶液中直至下沉
(4) 切片:脑组织冰冻切片20微米,37度烘干2h以上;
(5)将切片放置-80度保存,随用随取。
2免疫单标记
(1)做抗原修复(0.4g柠檬酸、3g柠檬酸钠、1000ml水配制成溶液,放到水浴箱中90~94℃保存15min),冷却后, 用1*PBS反复洗标本10min,吸去PBS,同时向标本加渗透液(配0.4%trition液,然后将BSA加入到配好的trition液中,配成1%浓度)100ul,常温下湿盒反应1h。
(2) 吸去渗透液,同时向标本加block液(5%封闭用正常山羊血清)100ul,常温下湿盒反应5h。
(3) 吸去block液,同时向标本加入1:50稀释的羊抗鼠一抗100ul,37度湿盒反应过夜。
(4)吸去一抗,并回收,用1*PBS洗液反复洗15min。
(5)快速从冰箱中取出驴抗羊的二抗,并立即用锡纸包裹,注入到一个新的以锡纸包裹的tube管中并用PBS以1:200稀释。
(6)进入暗室,吸去PBS,同时加入二抗100ul,37度湿盒反应2h,封片。
(7)放置到荧光显微镜下观察。
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.

