About InVivoMAb anti-mouse/human/rat v-H-Ras
The Y13-238 monoclonal antibody reacts with human, mouse, and rat v-H-ras (within amino acids 120-138) and does not react with v-K-ras. v-H-ras binds GTP/GDP and has intrinsic GTPase activity. Ras proteins alternate between an inactive form bound to GDP and an active form bound to GTP, activated by a guanine nucleotide-exchange factor (GEF) and inactivated by a GTPase-activating protein (GAP). Under normal conditions, Ras family members influence cell growth and differentiation events. Mutations in the Ras family of proto-oncogenes are very common, being found in 20% to 30% of all human tumors. Ras point mutations are the single most common abnormality of human proto-oncogenes. The Y13-238 antibody does not neutralize ras GTPase binding and hydrolysis in vivo or in vitro.
InVivoMAb anti-mouse/human/rat v-H-Ras Specifications
| Isotype | Rat IgG2a |
| Recommended Isotype Control(s) | InVivoMAb rat IgG2a isotype control, anti-trinitrophenol(BE0089) |
| Recommended InVivoPure Dilution Buffer | InVivoPure pH 7.0 Dilution Buffer(IP0070) |
| Immunogen | Harvey murine sarcoma virus infected NRK cells |
| Reported Applications | Immunoprecipitation |
| 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 G |
| Storage | The antibody solution should be stored at the stock concentration at 4°C. Do not freeze. |
| RRID | AB_10951512 |
| Molecular Weight | 150 kDa |
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英 ['aisəutaip]
美 ['aisəutaip]
n. 同形像;
同形像统计图
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?
阴性对照(Isotype Control):非特异荧光的强弱取决于抗体浓度、单克隆荧光抗体特异性和纯度,应与试验管抗体相对应。在多色分析时,同型对照应与其它抗体同时使用,以避免补偿造成的误差
血小板体外活化试验:使用正常人活化标本作为阳性质控;使用正常人未活化标本作为阴性质控
血小板自身抗体检测:使用含有已知血小板抗体的血清与血小板孵育,作为阳性质控;使用不含血小板抗体的血清与血小板孵育,作为阴性质控
血小板表面抗原缺失:如巨血小板症血小板表面CD42a/CD42b缺失,血小板无力症血小板表面gpIIb/IIIa,即CD41/CD61缺失或异常。使用正常人标本做阳性对照,抗体的同型对照做阴性对照
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。

