Anti-Phospho-JAK2 (Tyr1007, 1008) Polyclonal Antibody is a Rabbit antibody targeting Phospho-JAK2 (Tyr1007, 1008). Anti-Phospho-JAK2 (Tyr1007, 1008) Polyclonal Antibody can be used in IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

Human,Mouse,Rat,Zebrafish (predicted:Chicken,Pig,Rabbit)

1. From 《Cancer Medicine》(2016.6): PublitionDirect effect of dasatinib on signal transduction pathways associated with a rapid mobilization of cytotoxic lymphocytes , IF:2.5
Author: Noriyoshi Iriyama, Yoshihiro Hatta & Masami Takei
Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, Tokyo, Japan
2. Blank control: HL-60
Primary Antibody (green line): Rabbit Anti-phospho-JAK2 (Tyr1007+Tyr1008) antibody (TMAB-01440)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at-20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. Paraformaldehyde-fixed, paraffin embedded (rat lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (mouse lung); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (5 μg/ml, blue) was used to stain the cell nucleus.
7. Tissue/cell: HeLa cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (5 μg/ml, blue) was used to stain the cell nucleus.
8. Tissue/cell: NIH/3T3 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (5 μg/ml, blue) was used to stain the cell nucleus.
9. Tissue/cell: A549 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (phospho-JAK2 (Tyr1007+Tyr1008)) Polyclonal Antibody, Unconjugated (TMAB-01440) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (5 μg/ml, blue) was used to stain the cell nucleus.
10. Blank control (Black line): Raji (Black).
Primary Antibody (green line): Rabbit Anti-phospho-JAK2 (Tyr1007+Tyr1008) antibody (TMAB-01440)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA （10 min) and then permeabilized with 90% ice-cold methanol for 20 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human JAK2 around the phosphorylation site of Tyr1007/1008

Human

JAK2

Tyrosine-protein kinase JAK2

Cell cycle inhibitors,Kinases/phosphatases,Signal transducers,Cell Cycle Inhibitors,Kinases/Phosphatases,STATs,SARS Coronavirus,Tyrosine Kinases

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. 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.