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Description
Details
Description:Mouse monoclonal antibody to human interleukin 8 (IL-8)
Purification: Protein G affinity purified
Product Type:Primary antibody
Target Protein:Human IL-8
Immunogen:Purified recombinant human IL-8
Fusion Myeloma:P3X63Ag8.653
Specificity:This antibody recognizes both recombinant and native human IL-8.
Species Reactivity: Human, others not tested.
Cross-reactivity:This antibody does not cross react with human Monocyte Chemotactic Activating Factor (MCAF) or RANTES (Regulated on Activation, Normal T-cell Expressed, and Secreted).
Host / Isotype: Mouse, IgG1 Kappa
Formulation: Lyophilized in 0.01M PBS, pH 7.0.
Reconstitution:Double distilled water is recommended and to adjust the final concentration to 1.00 mg/mL.
Storage: Store at -20oC
Research Area:Cytokine, interleukin and chemotaxis
Background:
Interleukin 8 (IL-8), like IL-6, is secreted by macrophages and a variety of cells that express Toll-like receptors in response to the stimulation of pathogens. IL-8"s primary function is to recruit neutrophils and other target cells through chemotaxis to the infected site to eliminate pathogens. IL-8 causes increased intracellular Ca2+, release of reactive oxygen species, and other physiological changes required for migration and phagocytosis. IL-8 is also known to promote angiogenesis.
Applications:
ELISA: React with human IL-8.
Neutralizing: In chemotaxis assay, the antibody inhibited 97 % of the chemotactic activity of IL-8 on RB/293 cell using 5mg/ml of MAb and 10ng/ml of human IL-8 in assay. This antibody inhibited the chemotactic effect of human IL-8, and had no inhibitory effect to MIP 1-bata and RANTES in neutrophil chemotaxis assay, indicating that the neutralizing activity is specific. The antibody also neutralized IL-8 induced calcium influx in human granulocytes.
Western Blot: The antibody when used at 0.02-0.1mg/ml concentration, will allow visualization of 100ng/lane of recombinant human IL-8. Figure (see image): Western blot analysis of recombinant human IL-8 using clone I8-61.
References:
If research is published using this product, please inform Anogen in order to cite the reference on this datasheet. Anogen will provide one unit of product in the same category as gratitude.
Additional
Additional Information
Product Specificity | mAb anti-Human IL-8, I8-61 |
---|---|
Application | EIA, NT, WB |
Size | 0.1 mg |
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建立稳定细胞株,一般是根据不同基因载体中所含有的抗性标志选用相应的药物对靶...专注整体实验·服务生命科学已开展了数千个实验外包项目
一种是脂质体转染后,单克隆筛选稳定细胞株.另外一种是应用逆转录病毒,慢病毒转染,筛选稳定细胞株.
脂质体转染:在转染24小时后,消化细胞并计数.将细胞种到96孔板,保证每个孔2-3个细胞,这样才能得到单克隆.待细胞贴壁后,加入抗生素筛选.筛选时间和浓度视细胞而定.一般G418一个星期作用,嘌呤霉素2-3天.
脂质体法筛单克隆时间较长,且效率低,大概只有1%.
病毒转染:先要用包装细胞,一般为293细胞,包装出病毒,再用病毒转染目的细胞.包装病毒视不同类型的病毒而定,一般要3-5天的时间.包装好的病毒要测滴度,根据滴度决定转染目的细胞的病毒量.转染目的细胞1-2天后加抗生素筛选得到稳定细胞株.
病毒转染得到稳定细胞株的效率高,只是步骤繁琐.