Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP/山羊抗小鼠IgG (H+L)Superclonal™ 重组二抗, HRP

货号：A28177,A28177SAMPLE

规格：1mg,0.05mg

价格：2547,452

产品类型：荧光二抗

品牌：Thermo Fisher

抗原：Recombinant full-length protein

物种：小鼠

宿主：山羊

抗体亚型：IgG

荧光染料：HRP

抗体类型:	荧光二抗	同型对照：
浓度：	1 mg/mL	用法：	0.05-1 µg/mL（ELISA）； 1:10,000-1:200,000 (WB)

产品详细信息The sensitivity and specificity of each lot is confirmed using ELISA.Minimal cross-reactivity with rabbit, rat, human, bovine, guinea pig and donkey IgG is observed.Recombinant antibodies are produced using specific genes that code for the desired antibodies. These genes are cloned into an expression vector and expressed in vitro. The advantages of recombinant antibodies include: better specificity, animal origin-free formulation, and more lot-to-lot consistency. 靶标信息Anti-Mouse secondary antibodies are affinity-purified antibodies with well-characterized specificity for mouse immunoglobulins and are useful in the detection, sorting or purification of its specified target. Secondary antibodies offer increased versatility enabling users to use many detection systems (e.g. HRP, AP, fluorescence). They can also provide greater sensitivity through signal amplification as multiple secondary antibodies can bind to a single primary antibody. Most commonly, secondary antibodies are generated by immunizing the host animal with a pooled population of immunoglobulins from the target species and can be further purified and modified (i.e. immunoaffinity chromatography, antibody fragmentation, label conjugation, etc.) to generate highly specific reagents.

数据：

Mouse IgG (H+L) Secondary Antibody (A28177) in WB Western blot analysis of Phosphorylated Progesterone Receptor (upper panel) and total Progesterone Receptor (lower panel) was performed by loading 20 µg of nuclear (N) or cytoplasmic (C) T47D cell lysates, untreated (-) or stimulated (+) with 100 nm promegestone (R5020) for 1 hour and 10 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. Phosphorylated Progesterone Receptor was detected using a Phospho-Progesterone Receptor (Ser190) mouse monoclonal antibody (upper panel, Product # 37-8200) at a dilution of 1:1000 and Total Progesterone Receptor was detected using a Progesterone Receptor mouse monoclonal antibody (lower panel, Product # MA1-12626) at a concentration of 1 µg/mL in blocking buffer overnight at 4°C on a rocking platform, followed by a Superclonal goat anti-Mouse IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product # 62236). Mouse IgG (H+L) Secondary Antibody (A28177) in WB Western blot analysis of phospho-Histone H3 pSer10 was performed by loading 10 µg of acid extracted HeLa nuclear lysate extracted from cells not treated (lane 1) or treated with 100 nM calyculin A (Product # PHZ1044) (lane 2) for 30 min in reducing sample buffer (Product # 39000) and Page Ruler Plus Protein Ladder (Product # 26619) onto a Novex 4-20% Tris-Glycine polyacrylamide gel (Product # WT4201BX10). Proteins were transferred to nitrocellulose membrane (Product # 88018) with Transfer Buffer (Product # 84731) using the G2 Fast Blotter (Product # 62288). Membrane was blocked in StartingBlock T20 (Product # 37543) for 30 min at room temperature. Phospho-Histone H3 pSer10 was detected at approximately 17 kDa using an H3S10ph monoclonal antibody (Product # MA3-057) at a dilution of 1:2000 in StartingBlock T20 overnight at at 4°C on a rocking platform, followed by a goat anti-mouse superclonal IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:5000 for one hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product # 62236). _x000D_ Mouse IgG (H+L) Secondary Antibody (A28177) in WBWestern blot analysis was performed on whole cell extracts (30 µg lysate) of U-87 MG (Lane 1) and K-562 (Lane 2). The blots were probed with Anti SOD2 Mouse Monoclonal Antibody (Product# MA1-106, 0.25 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A28177) at dilutions 1:10,000 (Fig. 1), 1:100,000 (Fig. 2) and 1:200,000 (Fig. 3). A 25 kDa band corresponding to SOD2 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary antibody after blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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参考文献：

1.AutophagyElevated Mirc1/Mir17-92 cluster expression negatively regulates autophagy and CFTR (cystic fibrosis transmembrane conductance regulator) function in CF macrophages.

技术参数

产品应用 ELISA;WB

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