Mouse anti-Human IgE Secondary Antibody/小鼠抗人lgE二抗

货号：MII0201

规格：1 mg

价格：7669

产品类型：二抗

品牌：Thermo Fisher

抗原：Human Myeloma IgE

物种：人

宿主：小鼠

抗体亚型：IgE

克隆号：F4-#16

荧光染料：其它

类型:	二抗	同型对照：
浓度：	3.1 mg/mL	用法：	Assay Dependent(ELISA)；Assay Dependent(IP)；Assay Dependent(RIA) ；1:500-1:2000(WB)

产品详细信息MII0201 was formerly sold as a Seradyn product. This is a larger pack size of MA5-14704.靶标信息Thermo Scientific Anti-Human secondary antibodies are affinity-purified antibodies with well-characterized specificity for human 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

Human IgE Secondary Antibody (MII0201) in WBWestern blot analysis of human IgE was performed by loading 0.5 µg of purified human IgE, IgM, IgA and IgG per well and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-glycine polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blots were blocked with 5% milk in TBST for at least 1 hour at room temperature. Human IgE was detected at ~75 kD using a human IgE monoclonal antibody (Product # MII0201) at a dilution of 1:1000 in blocking buffer overnight at 4C on a rocking platform, followed by an HRP-conjugated goat anti-mouse light chain secondary antibody at a dilution of 1:40,000 for at least 30 minutes at room temperature (left blot). To control for protein loading, a duplicate blot was probed with a mixture of goat anti-human IgG, IgM, and IgA (Product # 31128) and goat anti-human IgE (Product # A18801) antibodies at concentrations of 1 µg/mL, followed by a mouse anti-goat IgG-HRP secondary antibody (Product # 31400) at a dilution of 1:8000 (right blot). Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080) Human IgE Secondary Antibody (MII0201) in ELISADirect ELISA analysis of human IgE was performed by coating wells of a 96-well plate with 100 µL per well of human IgE, IgM, IgA, or IgG, diluted in carbonate/bicarbonate buffer (Product # 28382) at concentrations ranging from 5-5000 ng/mL, and incubating overnight at 4C. The plate was washed and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543) for at least 30 minutes at room temperature, and then incubated with 100 µL per well of a human IgE monoclonal antibody (Product # MII0201) at a concentration of 1 µg/mL for 1 hour at room temperature. The plate was washed, and incubated with 100 µL per well of an HRP-conjugated goat anti-mouse IgG Fc secondary antibody (Product # 31439) at a dilution of 1:8000 for at least 30 minutes at room temperature. Detection was performed by adding 100 µL per well of 1-Step Ultra TMB substrate (Product # 34028) and incubating for a few minutes at room temperature. The reactions were then stopped with TMB Stop Solution (Product # N600). Absorbances were read on a spectrophotometer at 450-550 nm.

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