Product Name | NFkB-p65(Phospho-Thr254) Antibody |
Host Species | Rabbit |
Clonality | Polyclonal |
Purification | Antibodies were produced by immunizing rabbits with synthetic phosphopeptide and KLH conjugates. Antibodies were purified by affinity-chromatography using epitope-specific phosphopeptide. Non-phospho specific antibodies were removed by chromatogramphy using non-phosphopeptide. |
Applications | WB IHC IF |
Reactivity | Hu Ms Rt |
Specificity | The antibody detects endogenous level of NF-kB p65 only when phosphorylated at threonine 254. |
Immunogen | Peptide-KLH. Peptide sequence around phosphorylation site of threonine 254 (F-R-T(p)-P-P) derived from Human NFkB-p65. |
Target Name | NFkB-p65 |
Alternative Name | NFKB3; RELA; TF65; Transcription factor p65; p65 |
Modification | Phospho-Thr254 |
SDS-PAGE MW | 65kd |
Accession No | Q04206; NP_001138610.1 |
Concentration | 1.0mg/ml |
Formulation | Supplied at 1.0mg/mL in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. |
Storage | Store at -20°C for long term preservation (recommended). Store at 4°C for a short term use. |
Application Western blotting: 1:500~1:1000 Immunohistochemistry: 1:50~1:100 Immunofluorescence: 1:100~1:200
Background
NF-kappa-B is a pleiotropic transcription factor which is present in almost all cell types and is involved in many biological processed such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52 and the heterodimeric p65-p50 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity. Different dimer combinations act as transcriptional activators or repressors, respectively. NF-kappa-B is controlled by various mechanisms of post-translational modification and subcellular compartmentalization as well as by interactions with other cofactors or corepressors. NF-kappa-B complexes are held in the cytoplasm in an inactive state complexed with members of the NF-kappa-B inhibitor (I-kappa-B) family. In a conventional activation pathway, I-kappa-B is phosphorylated by I-kappa-B kinases (IKKs) in response to different activators, subsequently degraded thus liberating the active NF-kappa-B complex which translocates to the nucleus. NF-kappa-B heterodimeric p65-p50 and p65-c-Rel complexes are transcriptional activators. The NF-kappa-B p65-p65 complex appears to be involved in invasin-mediated activation of IL-8 expression. The inhibitory effect of I-kappa-B upon NF-kappa-B the cytoplasm is exerted primarily through the interaction with p65. p65 shows a weak DNA-binding site which could contribute directly to DNA binding in the NF-kappa-B complex.
Yeh PY, et al. (2004) J Biol Chem. 279(25): 26143-26148. Ryo A, et al. (2003) Mol Cell.12 (6): 1413-1426. Baeuerle P A, et al. (1994) Annu Rev Immunol. 12:141-179. Baeuerle P A, et al. (1996) Cell 87:13-20. Haskill S, et al. (1991) Cell 65:1281-1289.
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没有这个功能的机器，必须使用counting beads，也就是在一定体积的血制品样本中加入一定量已知体积的计数微球。这样微球的浓度已知，通过计数1000个微球，得到计数的细胞数量，从而计算出原来样品的细胞浓度。
变阻法计数在大多数细胞计数器中是利用小孔管换能器装置实现的。
在仪器的取样杯内装有一根吸样管，吸样管下部开有一个小孔(宝石制作)，因此也叫做小孔管。小孔管内外各置一只铂金电极，两电极间施加一个恒定的电流。测试时，先将待测血液用洁净的电解液充分稀释，使血细胞在电解液中成为游散状态，然后在小孔管上端施以负压，在负压的抽吸下，混有血细胞的电解液便被均匀地抽进小孔管。当血细胞通过小孔时，排开了等体积的电解液，使电解液的等效电阻瞬间变大，这个变大的电阻在恒流源的作用下引起一个等比例增大的电压。当细胞离开小孔附近后，电解液的等效阻值又恢复正常，直到下一个细胞到达小孔。这样血细胞连续地通过小孔，就在电极两端产生一连串电压脉冲。脉冲的个数与通过小孔的细胞个数相当，脉冲的幅度与细胞体积成正比。
需要测量转染之后细胞某mRNA表达变化。
实验组给予转染，空白组做对照。
做PCR的时候肯定是要求两组的细胞计数是一样的，请问是转染之前还是转染之后对两组进行计数呢？如果样品两边计数不一样，怎么让做实验的时候两边细胞数目变成一样呢？
这样的回答可能没分，好和不好都是比较出来的，一群国产的放在一起比总有个性能比较均衡，比较突出性价比好的，
还是没有和说那个厂家的好桂林的呢好像是优利特这个厂家在做，特康的前身呢就是百特，现在的百特呢基本上是在做试剂，这些东西呢。
也搜索的到的，不过答案还是比较接近的，但想知道的是国产那个厂家生产的比较好，而只回答了一半就和介绍了下国产的血球分析仪有那个几个大的厂家，没有和区分那个最好那个第二那个次点，或许一堆鸡蛋里实在挑不出骨头。
具所知呢迈瑞的好点特康实力没有迈瑞好，迈瑞吸引了一批外资在搞研发，桂林的呢好像有个优利特现在出血分析仪，他做的尿分析仪市场的占有还可以，血的反应不是很强烈。
实在是想找个内行的给比较下。
不用太精确的话计数板就可以了
过流式的话细胞里面要有稳定表达的荧光蛋白才行
2，细胞溶液必须均匀，再用之前要震荡均匀，否则细胞沉在试管底部，容易出现误差
