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JAK2 ELISA Kit (Rat)

CAT: 0247-OKCD02588-96WSize: 96 WellDry Ice: NoHazardous: No
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CAT#:0247-OKCD02588-96WSize:96 Well
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Reproducibility
Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Janus Kinase 2 (JAK2) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Janus Kinase 2 (JAK2) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%
CAS Number
7732-18-5
Gene Name
Janus kinase 2
Gene Aliases
JAK-2; Janus kinase 2; Janus kinase 2 (a protein tyrosine kinase) ; tyrosine-protein kinase JAK2.
Gene ID
24514
Accession Number
NP_113702.1
Reactivity
Rat|Rattus norvegicus
Target
Non-receptor tyrosine kinase involved in various processes such as cell growth, development, differentiation or histone modifications. Mediates essential signaling events in both innate and adaptive immunity. In the cytoplasm, plays a pivotal role in signal transduction via its association with type I receptors such as growth hormone (GHR), prolactin (PRLR), leptin (LEPR), erythropoietin (EPOR), thrombopoietin (THPO) ; or type II receptors including IFN-alpha, IFN-beta, IFN-gamma and multiple interleukins. Following ligand-binding to cell surface receptors, phosphorylates specific tyrosine residues on the cytoplasmic tails of the receptor, creating docking sites for STATs proteins. Subsequently, phosphorylates the STATs proteins once they are recruited to the receptor. Phosphorylated STATs then form homodimer or heterodimers and translocate to the nucleus to activate gene transcription. For example, cell stimulation with erythropoietin (EPO) during erythropoiesis leads to JAK2 autophosphorylation, activation, and its association with erythropoietin receptor (EPOR) that becomes phosphorylated in its cytoplasmic domain. Then, STAT5 (STAT5A or STAT5B) is recruited, phosphorylated and activated by JAK2. Once activated, dimerized STAT5 translocates into the nucleus and promotes the transcription of several essential genes involved in the modulation of erythropoiesis. Part of a signaling cascade that is activated by increased cellular retinol and that leads to the activation of STAT5 (STAT5A or STAT5B) . In addition, JAK2 mediates angiotensin-2-induced ARHGEF1 phosphorylation. Plays a role in cell cycle by phosphorylating CDKN1B. Cooperates with TEC through reciprocal phosphorylation to mediate cytokine-driven activation of FOS transcription. In the nucleus, plays a key role in chromatin by specifically mediating phosphorylation of 'Tyr-41' of histone H3 (H3Y41ph), a specific tag that promotes exclusion of CBX5 (HP1 alpha) from chromatin.
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 Janus Kinase 2 (JAK2) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Janus Kinase 2 (JAK2) . 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 Janus Kinase 2 (JAK2), 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 Janus Kinase 2 (JAK2) 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
0.055 ng/mL
Reconstitution
2°C to 8°C|-20°C
Components
Component Amount Anti-JAK2 Microplate 96 Wells (12 x 8 Well strips) JAK2 Lyophilized Standard 2 100X Biotinylated JAK2 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-JAK2 Microplate 96 Wells (12 x 8 Well strips) Anti-JAK2 Microplate 96 Wells (12 x 8 Well strips) JAK2 Lyophilized Standard 2 JAK2 Lyophilized Standard 2 100X Biotinylated JAK2 Detector Antibody 1 x 120 uL 100X Biotinylated JAK2 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 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 Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL
Specificity
This assay has high sensitivity and excellent specificity for detection of Janus Kinase 2 (JAK2) . No significant cross-reactivity or interference between Janus Kinase 2 (JAK2) and analogues was observed.
Protein Length
1132
NCBI Gene Symbol
Jak2
Protein Name
Tyrosine-protein kinase JAK2
Gene Name URL
Jak2
Nucleotide Accession Number
NM_031514.1