Rat PRL Antibody Pair Set
Price:
- 反应性： Rat
| Background |
Prolactin (PRL) is a neuroendocrine pituitary hormone. Prolactin is synthesized by the anterior pituitary,placenta,brain,uterus,dermal fibroblasts,decidua,B cells,T cells,NK cells and breast cancer cells. Originally characterized as a lactogenic hormone, further studies have demonstrated broader roles in breast cancer development,regulation of reproductive function,and immunoregulation. In the immune system,Prolactin has been shown to be secreted by human PBMC and to act as a proliferative growth factor. Additionally,Prolactin treatment of human PBMC has been shown to enhance IFN-gamma production. In the breast,Prolactin-induced morphogenesis of the mammary cells is mediated through IGF-2,which in turn upregulates cyclin D1. Prolactin has several molecular forms. The predominant form is a monomer,the non-glycosylated form is 23 kDa and the glycosylated form is 25 kDa. Glycosylated Prolactin is removed from the circulation faster and has been reported to have lower biological potency. Mouse Prolactin cDNA encodes a 228 amino acid (aa) residue protein with a putative 31 aa residue signal peptide. The Prolactin receptor is a transmembrane type I glycoprotein that belongs to the cytokine hematopoietic receptor family. B cells,T cells,macrophages,NK cells,monocytes,CD34+ progenitor cells,neutrophils,mammary gland,liver,kidney,adrenals,ovaries,testis,prostrate,seminal vesicles,and hypothalamus have all been shown to express the Prolactin receptor. Three forms of the receptor,generated by differential splicing,have been identified. These isoforms differ in the length of their cytoplasmic domains. It is believed that the short cytoplasmic form is non-functional. Prolactin signal transduction involves the JAK/STAT families and Src kinase family
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| Synonyms |
LTH;Lactotrope;Luteotropic Hormone
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| Swissprot | |
| Reactivity |
Rat
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| Specificity |
Detects Rat PRL in ELISAs
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| Buffer |
Capture Antibody:PBS with 0.04% Proclin 300, 50% glycerol, pH 7.4; Detection Antibody:PBS with 0.04% Proclin 300, 1% protective protein, 50% glycerol, pH 7.4
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| Storage |
-20℃
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| Expiration date |
12个月
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Q1:说明书中的标曲可以直接用来样本计算吗？
说明书上的标曲是指示标曲，主要是用于呈现拟合后标曲的形态以及最高/最低OD值的质控范围，不能直接使用。此外，受实验操作、实验环境以及仪器参数设定等因素的影响，实际操作获得的标曲OD值可能会跟说明书不完全一致（整体偏高或偏低），这种情况，建议通过在显色阶段控制前4个蓝色孔有明显颜色梯度，标曲最大OD值在1.2以上，以及标曲的相关系数在0.99以上，作为有效标曲。
Q2:如何进行超声操作？
使用超声波细胞破碎器超声处理悬浮液来裂解细胞（参考：用Soniprep150超声波发生器以振幅14μm超声处理30秒，破碎细胞；或者用超声破碎仪，200W，2秒/次，间隙3秒，总时间3-5分钟或者400A，5秒/次，间隙10秒，反复3-5次）。再于2-8℃，以1500×g离心10分钟，去除细胞碎片，收集上清液。超声过程中注意控制温度。
Q3:ELISA实验是否需要设置复孔？
为了保证实验结果的准确性，建议标准品和样品都做复孔检测。当然，ELISA实验并非要求一定做复孔/3复孔，客户可根据自己的需求设置实验的复孔与否。
Q4:稀释方案
稀释100倍：一步稀释。取5μl样本到495μl标准品&样本稀释液内，做100倍稀释； 稀释1000倍：两步稀释。取5μl样本到95μl标准品&样本稀释液内，做20倍稀释，再取5μl 20倍稀释样本到245μl标准品&样本稀释液内，做50倍稀释，总共稀释1000倍； 稀释100000倍：三步稀释。取5μl样本到195μl标准品&样本稀释液内，做40倍稀释，再取5μl 40倍稀释样本到245μl标准品&样本稀释液内，做50倍稀释，最后取5μl 2000倍稀释样本到245μl标准品&样本稀释液内，做50倍稀释，总共稀释100000倍； 每步稀释时取液量不少于3μl，稀释倍数不超过100倍。每步稀释都需混合均匀，避免起泡。
Q5:组织/细胞样本处理中提到的“反复冻融”过程该如何操作？
组织样本：组织研磨后，将其-80℃放置1小时/液氮放置0.5小时，然后置于30℃水浴锅中轻微振荡使其迅速融化，此操作反复1-2次即可。 细胞样本：按上述冻融的操作反复2-3次。若为膜蛋白可以适当超声一下，但需要控制好超声的温度和频率。 样本中建议提前加入蛋白酶抑制剂。常规推荐：PMSF(Cat.E-EL-SR002)。
Q6:ELISA试验是否需要技术重复？阳性或阴性对照如何设置？
建议客户做技术重复，一方面确认操作手法，一方面验证和提高实验结果的准确性。 ELISA实验中的阳性对照可以认为就是标准品。阴性对照是与待捡物具有同源性和同质性的空白基质或者已知浓度很低的基质溶液，一般较难获取，如果有可以设置为阴性对照。常规是设定空白对照，即样本稀释液。
