RAF1 (基因名), RAF proto-oncogene serine/threonine-protein kinase (蛋白名), raf1_human.
产品名称:
Human RAF1/ RAF proto-oncogene serine/threonine-protein kinase ELISA Kit
RAF原癌基因丝氨酸/苏氨酸蛋白激酶
货号:
E9583h
商标:
EIAab®
监管等级:
别名:
Proto-oncogene c-RAF, Raf-1, cRaf, RAF
检测方法:
ELISA
实验类型:
Sandwich
检测范围:
78-5000pg/mL
灵敏度:
20pg/mL
特异性:
Natural and recombinant human RAF proto-oncogene serine/threonine-protein kinase
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:
Cancer
通用注释
亚单元:
Monomer. Homodimer. Heterodimerizes with BRAF and this heterodimer possesses a highly increased kinase activity compared to the respective homodimers or monomers (PubMed:16508002). Heterodimerization is mitogen-regulated and enhanced by 14-3-3 proteins (PubMed:16508002). MAPK1/ERK2 activation can induce a negative feedback that promotes the dissociation of the heterodimer (PubMed:16508002). Forms a multiprotein complex with Ras (M-Ras/MRAS), SHOC2 and protein phosphatase 1 (PPP1CA, PPP1CB and PPP1CC) (PubMed:16630891). Interacts with Ras proteins; the interaction is antagonized by RIN1 (PubMed:11784866). Weakly interacts with RIT1. Interacts (via N-terminus) with RGS14 (via RBD domains); the interaction mediates the formation of a ternary complex with BRAF, a ternary complex inhibited by GNAI1 (By similarity). Interacts with STK3/MST2; the interaction inhibits its pro-apoptotic activity (PubMed:15618521). Interacts (when phosphorylated at Ser-259) with YWHAZ (unphosphorylated at 'Thr-232') (PubMed:9360956). Interacts with MAP2K1/MEK1 and MAP2K2/MEK2 (By similarity). Interacts with MAP3K5/ASF1 (via N-terminus) and this interaction inhibits the proapoptotic function of MAP3K5/ASK1 (PubMed:11427728). Interacts with PAK1 (via kinase domain) (PubMed:11733498). The phosphorylated form interacts with PIN1 (By similarity). The Ser-338 and Ser-339 phosphorylated form (by PAK1) interacts with BCL2 (PubMed:15849194). Interacts with PEBP1/RKIP and this interaction is enhanced if RAF1 is phosphorylated on residues Ser-338, Ser-339, Tyr-340 and Tyr-341 (PubMed:18294816). Interacts with ADCY2, ADCY5, ADCY6, DGKH, RCAN1/DSCR1, PPP1R12A, PKB/AKT1, PPP2CA, PPP2R1B, SPRY2, SPRY4, CNKSR1/CNK1, KSR2 and PHB/prohibitin (PubMed:10801873, PubMed:11719507, PubMed:12717443, PubMed:15385642, PubMed:15935327, PubMed:19710016, PubMed:10576742). Interacts with ROCK2 (By similarity). In its active form, interacts with PRMT5 (PubMed:21917714). Interacts with FAM83B; displaces 14-3-3 proteins from RAF1 and activates RAF1 (PubMed:22886302). Interacts with PDE8A; the interaction promotes RAF1 activity (PubMed:23509299).
功能:
Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases: ADCY2, ADCY5 and ADCY6, resulting in their activation. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Phosphorylates TNNT2/cardiac muscle troponin T. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
亚细胞位置:
Cytoplasm
Cell membrane
Mitochondrion
Nucleus
Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes. Phosphorylation at Ser-259 impairs its membrane accumulation. Recruited to the cell membrane by the active Ras protein. Phosphorylation at Ser-338 and Ser-339 by PAK1 is required for its mitochondrial localization. Retinoic acid-induced Ser-621 phosphorylated form of RAF1 is predominantly localized at the nucleus.
该产品尚未在任何出版物中被引用。
[1].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)ELISA试剂盒可以做多少个样本?
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)ELISA试剂盒使用视频?
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)ELISA试剂盒是放在-20℃冰箱保存吗?
EIAab的人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
如何分析人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
收集人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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?
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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].
人RAF原癌基因丝氨酸/苏氨酸蛋白激酶(RAF1)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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