Apex1 (基因名), DNA-(apurinic or apyrimidinic site) lyase (蛋白名), apex1_rat.
产品名称:
Rat Apex1/ DNA-(apurinic or apyrimidinic site) lyase ELISA Kit
DNA
货号:
E1939r
商标:
EIAab®
监管等级:
别名:
APEX nuclease, Apurinic-apyrimidinic endonuclease 1, REF-1, Redox factor-1, APEN, AP endonuclease 1, Ape, Apex, Ref1
检测方法:
ELISA
实验类型:
Sandwich
检测范围:
0.156-10ng/ml
灵敏度:
0.081ng/mL
特异性:
Natural and recombinant rat DNA-(apurinic or apyrimidinic site) lyase
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:
-
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: ≤4.5%
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: ≤7.4%
批内 CV: ≤4.5%
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: ≤7.4%
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 |
93 |
87-99 |
Plasma |
95 |
89-101 |
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) |
104-113% |
93-102% |
107-117% |
107-117% |
EDTA plasma(n=5) |
109-117% |
105-115% |
109-119% |
104-115% |
heparin plasma(n=5) |
85-97%
|
110-120% |
105-114% |
97-108% |
通用注释
亚单元:
Monomer. Homodimer; disulfide-linked. Component of the SET complex, composed of at least APEX1, SET, ANP32A, HMGB2, NME1 and TREX1. Associates with the dimer XRCC5/XRCC6 in a DNA-dependent manner. Interacts with SIRT1; the interaction is increased in the context of genotoxic stress. Interacts with HDAC1, HDAC2 and HDAC3; the interactions are not dependent on the APEX1 acetylation status. Interacts with XRCC1; the interaction is induced by SIRT1 and increased with the APEX1 acetylated form. Interacts with NPM1 (via N-terminal domain); the interaction is RNA-dependent and decreases in hydrogen peroxide-damaged cells. Interacts (via N-terminus) with YBX1 (via C-terminus); the interaction is increased in presence of APEX1 acetylated. Interacts with HNRNPL; the interaction is DNA-dependent. Interacts (via N-terminus) with KPNA1 and KPNA2. Interacts with TXN; the interaction stimulates the FOS/JUN AP-1 complex DNA-binding activity in a redox-dependent manner. Interacts with GZMA, KRT8, MDM2, POLB, PRDX6, PRPF19, RPLP0, TOMM20 and WDR77. Binds to CDK5.
功能:
Multifunctional protein that plays a central role in the cellular response to oxidative stress. The two major activities of APEX1 in DNA repair and redox regulation of transcriptional factors. Functions as a apurinic/apyrimidinic (AP) endodeoxyribonuclease in the DNA base excision repair (BER) pathway of DNA lesions induced by oxidative and alkylating agents. Initiates repair of AP sites in DNA by catalyzing hydrolytic incision of the phosphodiester backbone immediately adjacent to the damage, generating a single-strand break with 5'-deoxyribose phosphate and 3'-hydroxyl ends. Does also incise at AP sites in the DNA strand of DNA/RNA hybrids, single-stranded DNA regions of R-loop structures, and single-stranded RNA molecules. Has a 3'-5' exoribonuclease activity on mismatched deoxyribonucleotides at the 3' termini of nicked or gapped DNA molecules during short-patch BER. Possesses a DNA 3' phosphodiesterase activity capable of removing lesions (such as phosphoglycolate) blocking the 3' side of DNA strand breaks. May also play a role in the epigenetic regulation of gene expression by participating in DNA demethylation. Acts as a loading factor for POLB onto non-incised AP sites in DNA and stimulates the 5'-terminal deoxyribose 5'-phosphate (dRp) excision activity of POLB. Plays a role in the protection from granzymes-mediated cellular repair leading to cell death. Also involved in the DNA cleavage step of class switch recombination (CSR). On the other hand, APEX1 also exerts reversible nuclear redox activity to regulate DNA binding affinity and transcriptional activity of transcriptional factors by controlling the redox status of their DNA-binding domain, such as the FOS/JUN AP-1 complex after exposure to IR. Involved in calcium-dependent down-regulation of parathyroid hormone (PTH) expression by binding to negative calcium response elements (nCaREs). Together with HNRNPL or the dimer XRCC5/XRCC6, associates with nCaRE, acting as an activator of transcriptional repression. Stimulates the YBX1-mediated MDR1 promoter activity, when acetylated at Lys-6, leading to drug resistance. Acts also as an endoribonuclease involved in the control of single-stranded RNA metabolism. Plays a role in regulating MYC mRNA turnover by preferentially cleaving in between UA and CA dinucleotides of the MYC coding region determinant (CRD). In association with NMD1, plays a role in the rRNA quality control process during cell cycle progression. Associates, together with YBX1, on the MDR1 promoter. Together with NPM1, associates with rRNA. Binds DNA and RNA.
亚细胞位置:
DNA-(apurinic or apyrimidinic site) lyase, mitochondrial
Mitochondrion
The cleaved APEX2 is only detected in mitochondria. Translocation from the cytoplasm to the mitochondria is mediated by ROS signaling and cleavage mediated by granzyme A. Tom20-dependent translocated mitochondrial APEX1 level is significantly increased after genotoxic stress (By similarity).
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[1].
大鼠DNA(Apex1)ELISA试剂盒可以做多少个样本?
大鼠DNA(Apex1)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].
大鼠DNA(Apex1)ELISA试剂盒使用视频?
大鼠DNA(Apex1)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].
大鼠DNA(Apex1)ELISA试剂盒是放在-20℃冰箱保存吗?
EIAab的大鼠DNA(Apex1)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].
大鼠DNA(Apex1)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].
大鼠DNA(Apex1)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].
如何分析大鼠DNA(Apex1)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].
大鼠DNA(Apex1)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].
收集大鼠DNA(Apex1)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].
大鼠DNA(Apex1)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?
大鼠DNA(Apex1)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].
大鼠DNA(Apex1)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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