RAF1 ELISA Kit (Human)

• Reproducibility:

  Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level C-Raf Proto Oncogene Serine/Threonine Protein Kinase (Raf-1) 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 (Raf-1) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

• Gene Name:

  Raf-1 proto-oncogene, serine/threonine kinase

• Gene Aliases:

  CMD1NN; c-Raf; CRAF; C-Raf proto-oncogene, serine/threonine kinase; NS5; Oncogene RAF1; proto-oncogene c-RAF; raf proto-oncogene serine/threonine protein kinase; RAF proto-oncogene serine/threonine-protein kinase; Raf-1; v-raf-1 murine leukemia viral oncogene homolog 1; v-raf-1 murine leukemia viral oncogene-like protein 1.

• Gene ID:

  5894

• Accession Number:

  NP_002871

• Reactivity:

  Homo sapiens|Human

• Target:

  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.

• Type:

  ELISA Kit

• Applications:

  Enzyme-linked Immunosorbent assay-Sandwich

• Detection Method:

  Colorimetric

• Assay Principle:

  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 (Raf-1) . 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 (Raf-1) . 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 (Raf-1), 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 (Raf-1) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

• Assay Protocol:

  Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

• Sample Type:

  Tissue homogenates, cell lysates and other biological fluids

• Detection Range:

  0.156-10ng/mL

• Sensitivity:

  < 0.059 ng/mL

• Reconstitution:

  2°C to 8°C|-20°C

• Components:

  Component Amount Anti-RAF1 Microplate 96 Wells (12 x 8 Well Strips) RAF1 Lyophilized Standard 2 x 10 ng 100X Biotinylated RAF1 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-RAF1 Microplate 96 Wells (12 x 8 Well Strips) Anti-RAF1 Microplate 96 Wells (12 x 8 Well Strips) RAF1 Lyophilized Standard 2 x 10 ng RAF1 Lyophilized Standard 2 x 10 ng 100X Biotinylated RAF1 Detector Antibody 120 uL 100X Biotinylated RAF1 Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

• Notes:

  Formerly GWB-KBBVM3

• Specificity:

  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.

• Protein Length:

  648

• NCBI Gene Symbol:

  RAF1

• Protein Name:

  RAF proto-oncogene serine/threonine-protein kinase

• Gene Name URL:

  RAF1

• CAS Number:

  7732-18-5

• Nucleotide Accession Number:

  NM_002880.3