Sort By

STAT5b Polyclonal Antibody (Bethyl Laboratories)

STAT5b Polyclonal Antibody for Western Blot, IHC, IP

Phospho-STAT5 alpha (Tyr694) Polyclonal Antibody (Invitrogen™)

Phospho-STAT5 alpha (Tyr694) Polyclonal Antibody for Western Blot, IHC, ELISA

LanthaScreen™ STAT5 (JAK2 V617F) U2OS Cell Line

LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. The JAK2⁄STAT5 signaling pathway plays an essential role in blood cell formation in response to cytokines such as GM-CSF, IL-3, and EPO. The recent discovery of an activating mutation in JAK2 (V617F) present in high percentage of myeloproliferative disease (MPD) patients suggests that this mutant JAK2 activity is a potential therapeutic target for certain forms of MPD. The assay described in this summary makes use of a cell line engineered for expression of the constitutively-active mutant kinase JAK2 V617F. By co-expressing GFP-STAT5 in this background, the phosphorylation state of STAT5 (specifically residue Tyr 694⁄699) can be modulated with JAK2 V617F inhibitors and analyzed in cell lysates using an anti-STAT5 [pTyr 694⁄699] and LanthaScreenTM terbium-anti-mouse antibody pair. GFP-STAT5 Lentivirus was transduced into U2OS cells followed by selection with Blasticidin. The selected pool was then transfected with a GST-JAK2 V617F construct using LipofectamineTM LTX, followed by selection with Geneticin. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker, and has been screened for the constitutive expression of GFP-STAT5 and GST-JAK2 V617F. Using a lytic TR-FRET immuno-assay, this cell line is validated for IC50 and Z' under optimized conditions using JAK Inhibitor I (Pyridone 6) as a small molecule inhibitor for JAK2 V617F-mediated GFP-STAT5 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and lysis⁄equilibration time.

STAT5 alpha/beta Human InstantOne™ ELISA Kit (Invitrogen™)

STAT5 alpha/beta Human InstantOne™ ELISA Kit for ELISA

STAT5 alpha/beta (Phospho) [pY694/pY699] Multispecies InstantOne™ ELISA Kit (Invitrogen™)

STAT5 alpha/beta (Phospho) [pY694/pY699] Human InstantOne™ ELISA Kit for ELISA

Phospho-STAT5 beta (Tyr694) Monoclonal Antibody (ST5P-4A9) (Invitrogen™)

Phospho-STAT5 beta (Tyr694) Monoclonal Antibody for Western Blot, ELISA

STAT5 alpha/beta Monoclonal Antibody (ST5-8F7) (Invitrogen™)

STAT5 alpha/beta Monoclonal Antibody for Western Blot, IF, ICC, ELISA

STAT5 beta Antibody (13HCLC), ABfinity™ Rabbit Oligoclonal (Invitrogen™)

STAT5 beta Oligoclonal Antibody for Western Blot, ChIP

STAT5 beta Polyclonal Antibody (Invitrogen™)

STAT5 beta Polyclonal Antibody for Western Blot, IHC (P), IP, ELISA, GS

STAT5 beta Monoclonal Antibody (ST5b-10G1) (Invitrogen™)

STAT5 beta Monoclonal Antibody for Western Blot, IF, ICC, IHC (P), IP, ChIP, ELISA, GS

LanthaScreen™ STAT5 TF-1 Cell Line

LanthaScreen® GFP Cellular Assays allow for the analysis of post-translational modifications for a number of target proteins in an live-cell context. In hematopoeitic cells, the JAK2⁄STAT5 signaling pathway plays an essential role in blood cell formation in response to cytokines such as GM-CSF, IL-3, and EPO. In this pathway, binding of these cytokines to their respective cell surface receptors results in the activation of JAK2, which in turn phospho-activates STAT5 proteins at specific tyrosine residues (Tyr-694⁄699). LanthaScreenTM-STAT5 TF-1 is a human hematopoeitic cell line which constitutively expresses GFP-STAT5 fusion proteins. The JAK2⁄STAT5 signaling pathway is known to be functionally intact in this cell line, therefore the GFP-STAT5 fusion protein serves as a substrate for the inducible phosphorylation by JAK2. Using this cell line, a homogenous immuno-assay has been developed in which the phosphorylation state of GFP-STAT5 is detected in cell lysates using a LanthaScreenTM Terbium-anti-mouse and anti-phospho STAT5 [pTyr694⁄699] antibody pair, in a time-resolved FRET (TR-FRET) readout. GFP-STAT5 Lentivirus was transduced into TF-1 cells followed by selection with Blasticidine. This cell line is a clonal population isolated by flow cytometry using GFP fluorescence as sorting marker, and has been screened for the constitutive expression of GFP-STAT5 fusion protein. Using a lytic TR-FRET immuno-assay, this cell line is validated for EC50 and Z' under optimized conditions using GM-CSF as an agonist for JAK2 -mediated GFP-STAT5 phosphorylation. This assay has also been tested for assay performance under variable experimental conditions, including cell plating density, stimulation time, DMSO tolerance and lysis⁄equilibration time.

STAT5 beta Antibody (13H20L4), ABfinity™ Rabbit Monoclonal (Invitrogen™)

STAT5 beta Monoclonal Antibody for Western Blot, IF, ICC, IHC (P), ChIP

Phospho-STAT5 alpha/beta (Ser726, Ser731) Polyclonal Antibody (Invitrogen™)

Phospho-STAT5 alpha/beta (Ser726, Ser731) Polyclonal Antibody for Western Blot, IHC (P)

Phospho-STAT5 alpha/beta (Ser725, Ser730) Polyclonal Antibody (Invitrogen™)

Phospho-STAT5 alpha/beta (Ser725, Ser730) Polyclonal Antibody for Western Blot

STAT5 alpha/beta Polyclonal Antibody (Invitrogen™)

STAT5 alpha/beta Polyclonal Antibody for Western Blot, IHC (P)