ELISA/assay

C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF) ELISA Kit

MBS2020276

48-Strip-Wells

565 USD

ELISA Kit

C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF) ELISA Kit

C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF)

RAF1; Raf1; V-raf-1 Murine Leukemia Viral Oncogene Homolog 1; Proto-oncogene c-RAF

cRAF, partial; RAF proto-oncogene serine/threonine-protein kinase; RAF proto-oncogene serine/threonine-protein kinase; v-raf-leukemia viral oncogene 1; Proto-oncogene c-RAF; cRaf; Raf-1

CRAF

Raf1; Raf1; Craf1; Raf-1; c-Raf; v-Raf; AA990557; BB129353; 6430402F14Rik; D830050J10Rik; Craf; cRaf

RAF1_MOUSE

Human

97

This assay has high sensitivity and excellent specificity for detection of C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF). No significant cross-reactivity or interference between C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF) and analogues was observed.

For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 degree C upon receipt while the others should be at 4 degree C. For opened kit: When the kit is opened, the remaining reagents still need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and reseal along entire edge of zip-seal. The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Samples: Tissue homogenates, Cell lysates and other biological fluids. Assay Type: Sandwich. Detection Range: 0.156-10ng/mL. Sensitivity: Typically less than 0.059ng/mL.

Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100 . Intra-Assay: CV<10% . Inter-Assay: CV<12% . Assay Procedure Summary: 1. Prepare all reagents, samples and standards;. 2. Add 100uL standard or sample to each well. Incubate 2 hours at 37 degree C;. 3. Aspirate and add 100uL prepared Detection Reagent A. Incubate 1 hour at 37 degree C;. 4. Aspirate and wash 3 times;. 5. Add 100uL prepared Detection Reagent B. Incubate 30 minutes at 37 degree C;. 6. Aspirate and wash 5 times;. 7. Add 90uL Substrate Solution. Incubate 15-25 minutes at 37 degree C;. 8. Add 50uL Stop Solution. Read at 450nm immediately.

Principle of the Assay: The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm 10nm. The concentration of C-Raf Proto Oncogene Serine/Threonine Protein Kinase (CRAF) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

18150196

BAB83675.1

Q99N57

75,151 Da

ARMS-mediated Activation Pathway (1110440); Activation Of NMDA Receptor Upon Glutamate Binding And Postsynaptic Events Pathway (1111226); Acute Myeloid Leukemia Pathway (83310); Acute Myeloid Leukemia Pathway (529); Adaptive Immune System Pathway (1110669); Alcoholism Pathway (585577); Alcoholism Pathway (587116); Androgen Receptor Signaling Pathway (198319); Axon Guidance Pathway (1111058); B Cell Receptor Signaling Pathway (198285)

RAF1: a proto-oncogenic TKL kinase of the RAF family. Functions downstream of the Ras family of membrane associated GTPases to which it binds directly. Once activated Raf-1 can phosphorylate and activate MEK1/2, which in turn phosphorylate and activate ERK1/2. 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 (MEK1/2) and the extracellular signal-regulated kinases (ERK1/2). RAF1, subsequent to phosphorylation by PAK1, phosphorylates BAD (Bcl2-antagonist of cell death) at S75. 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 (ASK1 and MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation. Protein type: Oncoprotein; Protein kinase, TKL; Protein kinase, Ser/Thr (non-receptor); Kinase, protein; EC 2.7.11.1; TKL group; RAF family. Cellular Component: Golgi apparatus; mitochondrion; membrane; cytoplasm; plasma membrane; pseudopodium; cytosol; nucleus. Molecular Function: identical protein binding; mitogen-activated protein kinase kinase binding; metal ion binding; nucleotide binding; protein kinase activity; transferase activity; protein serine/threonine kinase activity; protein binding; Ras GTPase binding; MAP kinase kinase kinase activity; protein heterodimerization activity; transferase activity, transferring phosphorus-containing groups; receptor signaling protein activity; kinase activity; ATP binding. Biological Process: nerve growth factor receptor signaling pathway; activation of MAPKK activity; somatic stem cell maintenance; MAPKKK cascade; intermediate filament cytoskeleton organization and biogenesis; signal transduction; positive regulation of peptidyl-serine phosphorylation; protein amino acid phosphorylation; negative regulation of cell proliferation; response to hypoxia; cell glucose homeostasis; positive regulation of transcription from RNA polymerase II promoter; positive regulation of protein amino acid phosphorylation; cell differentiation; negative regulation of protein complex assembly; phosphorylation; negative regulation of apoptosis

48-Strip-Wells

565 USD

96-Strip-Wells

765

5x96-Strip-Wells

2975

10x96-Strip-Wells

5340