About InVivoMAb anti-mouse IL-1α
The ALF-161 monoclonal antibody reacts with precursor, secreted and membrane-associated forms of mouse IL-1α (interleukin 1 alpha) also known as lymphocyte activating factor (LAF), and mononuclear cell factor (MCF). IL-1α is a 17 kDa pro-inflammatory cytokine produced by a variety of cells, including macrophages, dendritic cells, T and B lymphocytes. IL-1α exerts a wide range of immune and inflammatory responses on a many cell types including lymphocytes, epithelial cells and fibroblasts. IL-1 is made up of IL-1α and IL-1β which are the products of distinct genes, but which are recognized by two distinct IL-1 receptors. The IL-1 receptor type I, a 80 kDa transmembrane protein with demonstrated IL-1 signaling function and the IL-1 receptor type II, a 68 kDa membrane protein with a relatively short cytoplasmic tail. The type II receptor acts as a decoy target for IL-1, inhibiting IL-1 activities by preventing the binding of IL-1 to the type I receptor. The ALF-161 antibody has been shown to neutralize the bioactivity of natural or recombinant IL-1α.
InVivoMAb anti-mouse IL-1α Specifications
| Isotype | Armenian Hamster IgG |
| Recommended Isotype Control(s) | InVivoMAbpolyclonal Armenian hamster IgG(BE0091) |
| Recommended InVivoPure Dilution Buffer | InVivoPure pH 7.0 Dilution Buffer(IP0070) |
| Immunogen | Recombinant mouse IL-1α |
| Reported Applications | in vivo IL-1α neutralization |
| Endotoxin |
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| Purity |
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| Formulation |
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| Sterility | 0.2 μM filtered |
| Production | Purified from tissue culture supernatant in an animal free facility |
| Purification | Protein A |
| Storage | The antibody solution should be stored at the stock concentration at 4°C. Do not freeze. |
| RRID | AB_2687724 |
| Molecular Weight | 150 kDa |
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英 ['aisəutaip]
美 ['aisəutaip]
n. 同形像;
同形像统计图
选用和特异性抗体同种亚型的,同样荧光素标记的抗体,采用同样的染色步骤染同样浓度的细胞。然后上机检测。如果细胞有非特异性的结合,那同型对照的荧光强度可能会比未染色的阴性对照要高一些。
同型对照只用于那种阳性和阴性之间没有明确分解的情况。如果阳性细胞群非常清楚,是不需要同型对照的。另外,在多色同染的情况下,也不采用同型对照,而是采用FMO(fluorensence minus one)对照。
另外,对于同型对照的使用,也有很多反对意见。就算是同型抗体,也可能会有不同的亲和力,并不一定就能真实反应非特异性的结合。
同型对照(阴性对照)管:A同型+B同型+C同型
单阳性对照
A检测管:A+B同型+C同型
B检测管:B+A同型+C同型
C检测管:C+A同型+B同型
不存在荧光重叠的两个抗体不需要调节补偿。
样本为PBMC,想用三色标记Treg细胞,分别为CD4-PECY,CD25-Fitc,Foxp3-PE,想知道如何设置荧光补偿,用的是BD的流式仪,1、是设置阴性管一个,单阳性管每种一个,样品管一个么?
荧光素强度依次为PE, PECY5 APC FITC, PE最强,FITC最弱。 所以,建议抗体组合为:
Foxp3-PE,CD4-FITC,CD25-PECy5.5或者CD25-APC。
同时购买三个相应的同型对照抗体。
样品管设置如下:
1. cell + FITC-isotype + Pecy5.5( or APC)-isotype + PE-isotype
2. Cell + FITC-isotype + CD25-mAb + PE-isotype
3. Cell + PE-mAb + FITC-isotype + Pecy5.5( or APC)-isotype
4. Cell + CD4-FITC + Foxp3-PE + CD25 -PECy5.5( or APC)。 [这一管的数据可以拿来分析用]。
在充分了解调整电压和补偿原理的基础上,也可以做一些简化。如果不太懂,就按照这个来。其中Foxp3的抗体和同型对照抗体都是在固定破膜以后再加,不是与CD4或者CD25同时加。
英 ['aɪsətaɪp]美 ['aɪsəˌtaɪp]
n. 同型动物(或植物),图形文字,象征性图像;同位型;同模;同号模式
其同型为;percp-cy5.5(25ug包装)浓度:0.2mg/ml,
那么如果CD45RA抗体如果加入20ul,那同型抗体应该加入多少ml:
我自己的初步计算是:1ul的3分之1.不知道我算对了吗?应该怎么加才好呢?

