
- Specification
- Product Description:
- This protein protein interaction antibody pair set comes with two antibodies to detect the protein-protein interaction, one against the AKT1 protein, and the other against the FOXO1 protein for use in in situ Proximity Ligation Assay. See Publication Reference below.
- Reactivity:
- Human
- Quality Control Testing:
- Protein protein interaction immunofluorescence result.
Representative image of Proximity Ligation Assay of protein-protein interactions between AKT1 and FOXO1. HeLa cells were stained with anti-AKT1 rabbit purified polyclonal antibody 1:1200 and anti-FOXO1 mouse monoclonal antibody 1:50. Each red dot represents the detection of protein-protein interaction complex. The images were analyzed using an optimized freeware (BlobFinder) download from The Centre for Image Analysis at Uppsala University.
- Supplied Product:
- Antibody pair set content: 1. AKT1 rabbit purified polyclonal antibody (100 ug) 2. FOXO1 mouse monoclonal antibody (40 ug) *Reagents are sufficient for at least 30-50 assays using recommended protocols.
- Storage Instruction:
- Store reagents of the antibody pair set at -20°C or lower. Please aliquot to avoid repeated freeze thaw cycle. Reagents should be returned to -20°C storage immediately after use.
- MSDS:
Download
- Datasheet:
Download
- Publication Reference
- 1.
- An analysis of protein-protein interactions in cross-talk pathways reveals CRKL as a novel prognostic marker in hepatocellular carcinoma.Liu CH, Chen TC, Chau GY, Jan YH, Chen CH, Hsu CN, Lin KT, Juang YL, Lu PJ, Cheng HC, Chen MH, Chang CF, Ting YS, Kao CY, Hsiao M, Huang CY. Mol Cell Proteomics. 2013 Feb 8. [Epub ahead of print]
- Applications
- In situ Proximity Ligation Assay (Cell)
- Application Image
- In situ Proximity Ligation Assay (Cell)
- AKT1
- FOXO1
- Gene Information
- Entrez GeneID:
- 207
- Gene Name:
- AKT1
- Gene Alias:
- AKT,MGC99656,PKB,PKB-ALPHA,PRKBA,RAC,RAC-ALPHA
- Gene Description:
- v-akt murine thymoma viral oncogene homolog 1
- Omim ID:
- 164730, 181500
- Gene Ontology:
- Hyperlink
- Gene Summary:
- The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq
- Other Designations:
- RAC-alpha serine/threonine-protein kinase,murine thymoma viral (v-akt) oncogene homolog-1,protein kinase B,rac protein kinase alpha
- Gene Information
- Entrez GeneID:
- 2308
- Gene Name:
- FOXO1
- Gene Alias:
- FKH1,FKHR,FOXO1A
- Gene Description:
- forkhead box O1
- Omim ID:
- 136533, 268220
- Gene Ontology:
- Hyperlink
- Gene Summary:
- This gene belongs to the forkhead family of transcription factors which are characterized by a distinct forkhead domain.The specific function of this gene has not yet been determined; however, it may play a role in myogenic growth and differentiation. Translocation of this gene with PAX3 has been associated with alveolar rhabdomyosarcoma. [provided by RefSeq
- Other Designations:
- OTTHUMP00000018301,forkhead box O1A (rhabdomyosarcoma),forkhead homolog in rhabdomyosarcoma,forkhead, Drosophila, homolog of, in rhabdomyosarcoma
- Interactome
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的抗体不容易染色,哪为同仁有这方面的经验?另外基因公司卖的cellsignalingtechnology的抗体要1875元,而中山公司卖的这个抗体只有800多,其中差距在哪里?请赐教
GlucosestarvationcausestranslocationofAMPKβ2tothelysosomeinHEK-293cellsthatisdependentonN-myristoylation.Theexperimentwasperformedinβ2KOcellsasinFig.1c,exceptthatthelysosomalMarkerLAMP1(taggedwithRFP)wasco-expressedwiththewild-typeormutantAMPKβ2.Upperpanelsshowmergedimagesstainedblue(4′,6-diamidino-2-phenylindole(DAPI),nuclei),red(LAMP1,lysosomes)andgreen(AMPKβ2,detectedusingantibodyvalidatedine),incellsincubatedwithorwithoutglucosefor20 min.Lowersmallpanelsaremagnificationsoftheareasindicatedbydashedboxesintheupperpanels,showing(LtoR)redandgreenchannelsandmergedimages.
下面的这段话是图注,图注的意思我明白,但是我想知道merge后的图看什么颜色的荧光,蓝色是细胞核,红色是lysosome(位于胞质),绿色是AMPKβ2,该实验是想观察AMPKβ2是否转位到lysosome上了,如果确实发生了AMPKβ2转位到lysosome上,那么merge后是红色与绿色融合在一起,是吗?融合在一起发什么颜色的光了?
1)做免疫组化的片子最好不要同时做HE染色,因为那样会掩盖阳性着色的结果;而且用苏木素复染细胞核也是淡染,若要在免疫组化的片子上观察形态学的改变,除非有非常典型的表现,一般是有很大的难度的。
2)你是培养的细胞做免疫组化吗?如果是的话,那就每个样本只有一张片子吧?唯一的办法是进行免疫双染,同时做CD34和AFP(甲胎蛋白)。我觉得后者阳性便可以或多或少的获得向肝细胞分化的证据。
3)用HE染色来识别造血干细胞向肝细胞分化,也就是说要看到肝细胞的典型形态学改变后才可以下结论。问题是:早期的肝细胞仅凭形细胞态学观察并不能很容易的被识别。病理大夫看组织切片,重要的是看其组织学结构,对肝细胞也是这样。如果单拿出来一个细胞,还是培养的细胞,染色后真的不容易区分。所以我觉得你设计的用HE染色来确定早期细胞向哪个方向分化不是很有说服力。
4)如果是切片,也就是说每个样本可以有多张切片,那可以在连续切片上分别染CD34和AFP,拍照时选择相同视野,也有一定说服力。但是还是不如双染法更可靠。
5)用双染最大的问题在于:这两者都表达在胞浆中,染色容易相互覆盖。所以在双染的方法选择上你还要再考虑一下,可不可以用免疫荧光双染?这样也很有说服力的
免疫组织化学又称免疫细胞化学,是指带显色剂标记的特异性抗体在组织细胞原位通过抗原抗体反应和组织化学的呈色反应,对相应抗原进行定性、定位、定量测定的一项新技术。它把免疫反应的特异性、组织化学的可见性巧妙地结合起来,借助显微镜的现像和放大作用,在细胞,亚细胞水平检测各种抗原物质,并可在原位显示相应的基因和基因表达产物。免疫组织化学技术现已有:免疫荧光组织(细胞)化学技术、免疫酶组织(细胞)化学技术、亲和组织化学技术、免疫金银及铁标记免疫组织化学技术等。免疫荧光组织(细胞)化学技术是采用荧光素标记的已知抗体(或抗原)作为探针,检测待测组织、细胞标本中的靶抗原(或抗体),形成的抗原抗体复合物上带有荧光素,在荧光显微镜下,由于受高压汞灯光源的紫外光照射,荧光素发出明亮的荧光,这样就可以分辨出抗原(或抗体)的所在位置及其性质,并可利用荧光定量技术计算抗原的含量。以达到对抗原物质定位、定性、定量测定的目的。

