Rela (基因名), Transcription factor p65 (蛋白名), tf65_mouse.
产品名称:
Mouse Rela/ Transcription factor p65 ELISA Kit
转录因子p65
货号:
E0616m
商标:
EIAab®
监管等级:
别名:
Nuclear factor NF-kappa-B p65 subunit, Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3, Nfkb3
检测方法:
ELISA
实验类型:
Sandwich
检测范围:
0.235-15ng/mL
灵敏度:
0.121ng/mL
特异性:
Natural and recombinant mouse Transcription factor p65
样品类型:
Serum, plasma, tissue homogenates, cell culture supernates and other biological fluids
样品数据:
登录.
实验步骤:
研究领域:
Neurosciences
精密度
批内差:已知浓度的3个样本在一个板子内重复检测20次,以评估批内精密度。
批内 CV: ≤8.3%
批间差:已知浓度的3个样本在不同的板子上重复测定5次,以评估测定批间精密度。
批间 CV: ≤9.6%
批内 CV: ≤8.3%
批间差:已知浓度的3个样本在不同的板子上重复测定5次,以评估测定批间精密度。
批间 CV: ≤9.6%
回收率
回收率:低、中和高浓度的分析物被掺入到血清或者血浆样本中,进行回收实验测定。
Sample Type |
Average(%) |
Recovery Range(%) |
Serum |
94 |
88-100 |
Plasma |
96 |
90-102 |
线性
线性:给定样本通过梯度稀释,每次稀释的测量值与理论值的比值。
Sample |
1:2 |
1:4 |
1:8 |
1:16 |
serum(n=5) |
93-103% |
92-105% |
89-97% |
102-110% |
EDTA plasma(n=5) |
97-106% |
80-92% |
104-116% |
103-112% |
heparin plasma(n=5) |
89-99%
|
100-110% |
93-103% |
94-103% |
通用注释
亚单元:
Component of the NF-kappa-B p65-p50 complex. Component of the NF-kappa-B p65-c-Rel complex. Homodimer; component of the NF-kappa-B p65-p65 complex. Component of the NF-kappa-B p65-p52 complex. May interact with ETHE1. Binds AES and TLE1. Interacts with TP53BP2. Binds to and is phosphorylated by the activated form of either RPS6KA4 or RPS6KA5. Interacts with ING4 and this interaction may be indirect. Interacts with CARM1, USP48 and UNC5CL. Interacts with IRAK1BP1. Interacts with NFKBIA. Interacts with GSK3B. Interacts with NFKBIB. Interacts with NFKBIE. Interacts with NFKBIZ. Part of a 70-90 kDa complex at least consisting of CHUK, IKBKB, NFKBIA, RELA, IKBKAP and MAP3K14. Interacts with HDAC3; HDAC3 mediates the deacetylation of RELA. Interacts with HDAC1; the interaction requires non-phosphorylated RELA. Interacts with CBP; the interaction requires phosphorylated RELA. Interacts (phosphorylated at 'Thr-254') with PIN1; the interaction inhibits p65 binding to NFKBIA. Interacts with SOCS1. Interacts with MTDH and PHF11. Interacts with NFKBID. Interacts with ARRB2. Interacts with NFKBIA (when phosphorylated), the interaction is direct; phosphorylated NFKBIA is associated with a SCF(BTRC)-like complex lacking CUL1. Interacts with RNF25. Interacts (via C-terminus) with DDX1 (By similarity). Interacts with UFL1 and COMMD1 (By similarity). Interacts with BRMS1; this promotes deacetylation of 'Lys-310'. Interacts (when acetylated at Lys-310) with BRD4; leading to activation of the NF-kappa-B pathway (By similarity). Interacts with EHMT1 (via ANK repeats). Interacts with NOTCH2. Directly interacts with MEN1; this interaction represses NFKB-mediated transactivation (By similarity). Interacts with AKIP1, which promotes the phosphorylation and nuclear retention of RELA (By similarity). Interacts (via the RHD) with GFI1; the interaction, after bacterial lipopolysaccharide (LPS) stimulation, inhibits the transcriptional activity by interfering with the DNA-binding activity to target gene promoter DNA. Interacts with MEFV (By similarity). Interacts with CLOCK. Interacts with FOXP3 (By similarity). Interacts (via N-terminus) with CPEN1; this interaction induces proteolytic cleavage of p65/RELA subunit and inhibition of NF-kappa-B transcriptional activity (By similarity). Interacts with CDK5RAP3; stimulates the interaction of RELA with HDAC1, HDAC2 and HDAC3 thereby inhibiting NF-kappa-B transcriptional activity.
功能:
NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes 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 (By similarity). 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. Associates with chromatin at the NF-kappa-B promoter region via association with DDX1. Essential for cytokine gene expression in T-cells.
亚细胞位置:
Nucleus
Cytoplasm
Colocalized with DDX1 in the nucleus upon TNF-alpha induction (By similarity). Nuclear, but also found in the cytoplasm in an inactive form complexed to an inhibitor (I-kappa-B). Colocalizes with GFI1 in the nucleus after lipopolysaccharide (LPS) stimulation.
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[1].
小鼠转录因子p65(Rela)ELISA试剂盒可以做多少个样本?
小鼠转录因子p65(Rela)ELISA试剂盒分为2种规格,96孔和48孔。96孔的试剂盒,标曲和样本都做复孔的话,可以检测40个样本。96孔的试剂盒,标曲和样本都不做复孔的话,可以检测88个样本。
[2].
小鼠转录因子p65(Rela)ELISA试剂盒使用视频?
小鼠转录因子p65(Rela)ELISA试剂盒实验操作视频在以下网址中,对每一步的实验步骤都做了演示,方便实验员能更好地理解ELISA实验的过程。
https://www.eiaab.com.cn/lesson-tech/805.html
https://www.eiaab.com.cn/lesson-tech/805.html
[3].
小鼠转录因子p65(Rela)ELISA试剂盒是放在-20℃冰箱保存吗?
EIAab的小鼠转录因子p65(Rela)ELISA试剂盒,洗涤液、底物、终止液保存于4℃,其余试剂-20℃冰箱保存。
[4].
小鼠转录因子p65(Rela)ELISA试剂盒原理?
双抗体夹心法:用纯化的抗体包被微孔板,制成固相抗体,往包被有固相抗体的微孔中依次加入标准品或受检样本、生物素化抗体、HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
竞争法:用纯化的抗体包被微孔板,制成固相抗体,往包被有固相抗体的微孔中依次加入标准品或受检样本和生物素标记的目标分析物,受检标本中抗原与生物素标记抗原竞争结合有限的抗体。再加入HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
竞争法:用纯化的抗体包被微孔板,制成固相抗体,往包被有固相抗体的微孔中依次加入标准品或受检样本和生物素标记的目标分析物,受检标本中抗原与生物素标记抗原竞争结合有限的抗体。再加入HRP标记的亲和素,经过彻底洗涤后用底物TMB显色。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
[5].
小鼠转录因子p65(Rela)ELISA试剂盒中需要使用的样品量是多少?
夹心法100μL/孔,竞争法50μL/孔。如样本浓度过高时,应对样本进行稀释,以使稀释后的样本符合试剂盒的检测范围,计算时再乘以相应的稀释倍数。
[6].
如何分析小鼠转录因子p65(Rela)ELISA试剂盒数据?
建议标准曲线,并计算样本浓度。对于elisa的曲线拟合,一般建议采用4参数曲线拟合,4参数曲线拟合通常更适合免疫分析。推荐使用专业软件进行曲线拟合,例如curve expert 1.3。根据样本的OD值由标曲查出相应的浓度,再乘以稀释倍数;或用标准物的浓度与OD值计算出标曲的回归方程式,将样本的OD值代入方程式,计算出样本浓度,再乘以稀释倍数,即为样本的实际浓度。以下链接是curve expert 1.3软件拟合曲线的方法。
https://www.eiaab.com.cn/news/502/
https://www.eiaab.com.cn/news/502/
[7].
小鼠转录因子p65(Rela)ELISA试剂盒中是否包含人和动物的副产物,是否包含感染的或者传染性原料如HIV等?
除了抗体和稀释液中的BSA,不含其它人和动物的副产物,也不含感染材料。
[8].
收集小鼠转录因子p65(Rela)ELISA试剂盒血浆样本,用什么作为抗凝剂?
一般建议用EDTA和肝素作为抗凝剂。
[9].
小鼠转录因子p65(Rela)ELISA试剂盒酶标板可以拆成几部分?拆的时候是否需要避光,无菌?
小鼠转录因子p65(Rela)ELISA试剂盒酶标板是8×12孔条,可拆卸,板子可以拆成12条,注意避免孔污染,不需要避光和无菌。暂时不用的板子,放回原来装的袋子里,密封保存。
[10].
小鼠转录因子p65(Rela)ELISA试剂盒样本如何保存?
尽量检测新鲜样本。若无新鲜样本,则4℃保存1周,-20℃保存1个月,-80℃保存2个月。
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