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Stat5

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STAT5 is a laboratory equipment product manufactured by BD. It is a tool used for the detection and analysis of proteins in biological samples. The core function of STAT5 is to enable researchers to measure the levels and activation state of the STAT5 protein, which plays a crucial role in cellular signaling pathways.

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Lab products found in correlation

β actin, by Merck Group (3 mentions) Ecl chemiluminescence detection system, by GE Healthcare (2 mentions) P y694 699 stat5, by Cell Signaling Technology (2 mentions) Alexa fluor 680 goat anti mouse, by Thermo Fisher Scientific (2 mentions) Hsp90, by BD (2 mentions) Ripk3, by Cell Signaling Technology (2 mentions) Phospho p70 s6 kinase, by Cell Signaling Technology (2 mentions) Phospho mek1 2, by Cell Signaling Technology (2 mentions) Erk1 2, by Merck Group (2 mentions) P70 s6 kinase, by Santa Cruz Biotechnology (2 mentions) Ha 7 hrp, by Merck Group (2 mentions) Phospho stat5a b, by Merck Group (2 mentions) Actin aan01 a, by Cytoskeleton (2 mentions) Goat anti mouse hrp, by Jackson ImmunoResearch (2 mentions) Phospho erk1 2, by Cell Signaling Technology (2 mentions) Mek1 2, by Cell Signaling Technology (2 mentions) Goat anti rabbit hrp, by Jackson ImmunoResearch (2 mentions) Gapdh, by Abcam (2 mentions) Catalase, by Cell Signaling Technology (1 mentions) Nitrocellulose membrane, by GE Healthcare (1 mentions)

Close spelling variants of this product

Phospho-STAT5 (7 citations) Anti-STAT5 (5 citations) Anti-STAT5-pY694 (5 citations) Anti-phospho STAT5 (PY694) (5 citations) (47/Stat5(pY694)) (4 citations) Alexa Fluor 647 Mouse Anti-Stat5 (pY694); (4 citations) Anti-phospho-STAT5 (2 citations) Alexa Fluor 647-anti-phospho-STAT5 (Tyr694) (clone 47/Stat5, (2 citations) PSTAT5 [47/Stat5(pY694)] (2 citations) Anti-pY-STAT5 antibody (2 citations)

9 protocols using stat5

1

Immunoblotting analysis of STAT5 phosphorylation

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Cells were suspended in Laemmli’s 2× buffer (Bio-Rad, Hercules, CA, USA), separated on SDS/PAGE and blotted onto nitrocellulose membrane. Blots were incubated with the following antibodies (Abs): P-Y694/699-STAT5, Actin (Cell Signaling Technology, Danvers, MA, USA), STAT5 (BD Transduction Laboratories, Franklin Lakes, NJ, USA), STAT5A and STAT5B (Zymed/ThermoFisher Scientific, Waltham, MA, USA). Membranes were developed with the ECL chemiluminescence detection system (GE Healthcare, Little Chalfont Buckinghamshire, UK) using specific peroxidase (HRP) conjugated to rabbit or mouse IgG antibodies (Cell Signaling Technology).
Brachet-Botineau M., Deynoux M., Vallet N., Polomski M., Juen L., Hérault O., Mazurier F., Viaud-Massuard M.C., Prié G, & Gouilleux F. (2019). A Novel Inhibitor of STAT5 Signaling Overcomes Chemotherapy Resistance in Myeloid Leukemia Cells. Cancers, 11(12), 2043.
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2

Protein Immunoblotting Protocol

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NP40 cell lysates (1% NP40, 10% glycerol, 0.05 M Tris pH 7.5, 0.15 M NaCl, 1 mM PMSF, protease and phosphatase Inhibitor cocktails) (Roche and Thermo Scientific, respectively) were resuspended in Laemmli's 2x buffer, separated on SDS/PAGE and blotted onto nitrocellulose membrane (GE Healthcare, Little Chalfont, United Kingdom). Blots were incubated with the following antibodies (Abs): P-Y694/699-STAT5, catalase (Cell Signaling Technology, Danvers, United States), STAT5 (BD Transduction Laboratories, Franklin Lakes, United States), actin (Santa Cruz, Dallas, United States), Flag M2 (Stratagene, Santa Clara, United States), Glutaredoxin1 (Glrx1) (R&D, Minneapolis, United States). Membranes were developed with the ECL chemiluminescence detection system (GE Healthcare, Little Chalfont, United Kingdom) using specific peroxidase (HRP) conjugated to rabbit or mouse IgG antibodies (Cell signaling Technology, Danvers, United States) or goat IgG (Santa Cruz, Dallas, United States).
Bourgeais J., Ishac N., Medrzycki M., Brachet-Botineau M., Desbourdes L., Gouilleux-Gruart V., Pecnard E., Rouleux-Bonnin F., Gyan E., Domenech J., Mazurier F., Moriggl R., Bunting K.D., Herault O, & Gouilleux F. (2016). Oncogenic STAT5 signaling promotes oxidative stress in chronic myeloid leukemia cells by repressing antioxidant defenses. Oncotarget, 8(26), 41876-41889.
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3

BCR-ABL1 Signaling Pathway Analysis

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Ba/F3 cells expressing native or mutant BCR-ABL1 or primary CML patient cells were cultured for 4–6 hr in standard medium alone or with escalating concentrations of TKI(s), followed by lysis and boiling for 10 min in SDS-PAGE loading buffer. Lysates were separated on 4–15% Tris-glycine gels, transferred, and immunoblotted with antibodies for the BCR N-terminus (3902; Cell Signaling), phospho-ABLl (Y393 [1a numbering]; 2865; Cell Signaling), ABL1 (554148; BD Biosciences), phospho-CRKL (Tyr207) (3181S; Cell Signaling), phospho-STAT5 (Tyr694) (9351S; Cell Signaling), STAT5 (610192; BD Biosciences), β-actin (2691430; Millipore) and/or β-tubulin (05–66; Millipore).
Eide C.A., Zabriskie M.S., Savage Stevens S.L., Antelope O., Vellore N.A., Than H., Schultz A.R., Clair P., Bowler A.D., Pomicter A.D., Yan D., Senina A.V., Qiang W., Kelley T.W., Szankasi P., Heinrich M.C., Tyner J.W., Rea D., Cayuela J.M., Kim D.W., Tognon C.E., O’Hare T., Druker B.J, & Deininger M.W. (2019). Combining the Allosteric Inhibitor Asciminib with Ponatinib Suppresses Emergence of and Restores Efficacy Against Highly Resistant BCR-ABL1 Mutants. Cancer cell, 36(4), 431-443.e5.
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4

Antibody Validation for Signaling Pathways

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Antibodies used were HA (SC-805, Santa Cruz, Dallas, TX), HA-7-HRP (H6533, Sigma-Aldrich), MEK1/2 (#9126, Cell Signaling, Danvers, MA), phospho-MEK1/2 (#2338, Cell Signaling), ERK1/2 (M5670, Sigma-Aldrich), phospho- ERK1/2 (#4370, Cell Signaling), STAT5 (610191, BD Biosciences, Franklin Lakes, NJ), phospho-STAT5A/B (05–886R, Merck Millipore), phospho-p70 S6 kinase (#9234, Cell Signaling), p70 S6 kinase (SC-230, Santa Cruz), RIPK3 (#12107, Cell Signaling), HSP90 (610418, BD Transduction Laboratories), actin (AAN01-A, Cytoskeleton, Denver, CO), and tubulin (ab7291, Abcam, Cambridge, UK). The secondary antibodies used were goat anti-mouse HRP (115–035-003, Jackson ImmunoResearch, West Grove, PA), goat anti-rabbit HRP (111–035-003, Jackson ImmunoResearch), and Alexa Fluor 680 goat anti-mouse (A-21057, Molecular probes, Grand Island, NY).
Bigenzahn J.W., Fauster A., Rebsamen M., Kandasamy R.K., Scorzoni S., Vladimer G.I., Müller A.C., Gstaiger M., Zuber J., Bennett K.L, & Superti-Furga G. (2015). An Inducible Retroviral Expression System for Tandem Affinity Purification Mass-Spectrometry-Based Proteomics Identifies Mixed Lineage Kinase Domain-like Protein (MLKL) as an Heat Shock Protein 90 (HSP90) Client. Molecular & Cellular Proteomics : MCP, 15(3), 1139-1150.
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5

Immunoblotting of STAT Proteins

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Total cell lysates were prepared in lysis buffer (20 mM Hepes, pH 7.4, 20 mM NaCl, 10% Glycerol, and 1% Triton X-100) and protease and phosphatase inhibitor cocktails (CalBiochem). Blots were probed with antibodies that are specific for p-STAT1 (Y701, BD Biosciences), STAT1 (BD Biosciences), STAT2 (Santa Cruz), STAT3 (BD Biosciences), STAT4 (Santa Cruz), STAT5 (BD Biosciences), STAT6 (BD Biosciences), GAPDH (Santa Cruz), and β-actin (sigma).
Xiao W., Klement J.D., Lu C., Ibrahim M.L, & Liu K. (2018). IFNAR1 controls autocrine type I interferon regulation of PD-L1 expression in myeloid-derived suppressor cells. Journal of immunology (Baltimore, Md. : 1950), 201(1), 264-277.
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6

Immunoblot Analysis of Cellular Proteins

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Immunoblots were performed according to [11 (link)]. For each membrane, a housekeeping protein is depicted (HSP90, GAPDH, vinculin), which is always shown underneath the detected proteins of interest. Antibodies were: BCL-XL (#ab32370), GAPDH (#ab128915), WT1 (#ab89901) from Abcam, Cambridge, U.K.; RAF1 (#sc-133), HSP90 (#sc-13119), vinculin (#sc-736), ɣH2AX (#sc-101696) from Santa Cruz, Heidelberg, Germany; cleaved caspase-3 (#cs9661), p-Tyr202/Tyr204-ERK1/ERK2 (#cs9101), ERK1/ERK2 (#cs9102) from Cell Signaling, Leiden, Netherlands; p-Tyr694-STAT5 (#MA5-14973) from Thermo Fisher, Braunschweig, Germany; STAT5 (#610192) from BD Bioscience, Heidelberg, Germany; and PARP1 (#556362) from Pharmingen, Heidelberg, Germany. The protein ladders used were the prestained ScientificTM PageRulerTM (#26617) and the PageRulerTM Plus (#26620) from Thermo Fisher, Braunschweig, Germany.
Pons M., Zeyn Y., Zahn S., Mahendrarajah N., Page B.D., Gunning P.T., Moriggl R., Brenner W., Butter F, & Krämer O.H. (2021). Oncogenic Kinase Cascades Induce Molecular Mechanisms That Protect Leukemic Cell Models from Lethal Effects of De Novo dNTP Synthesis Inhibition. Cancers, 13(14), 3464.
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7

Immunoblotting Antibody Panel for AML

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Primary antibodies used for immunoblotting included NOX4 (Ab109225; Abcam; used for human primary AML samples), NOX4 (NB110-58849; Novus Biologicals; used for all other analyses), p22phox (#SC20781; Santa Cruz Biotechnology), β-Actin (A5441), α-Tubulin (T5168) (Sigma), Vinculin (BZL03106; Biozol), NUP98 (Ab179911 (13C1)), Histone H3 (Ab1220), Calreticulin (Ab22683), KDEL (Ab12223) (Abcam), GAPDH (#5174), Lamin A/C (#2032), HDAC1 (#5356), pAKT (Ser473; #9271), AKT(#9272), pERK1/2 (Thr202/Tyr204; #9106), ERK1/2 (#4696), pGSK3β (Ser9; #9336), GSK3β (#9315) (Cell Signaling), pSTAT5 (Tyr694/699; #04-886; Millipore), STAT5 (#610191; BD Biosciences) and HA (#MMS-101R; Cambridge Bioscience).
Moloney J.N., Jayavelu A.K., Stanicka J., Roche S.L., O'Brien R.L., Scholl S., Böhmer F.D, & Cotter T.G. (2017). Nuclear membrane-localised NOX4D generates pro-survival ROS in FLT3-ITD-expressing AML. Oncotarget, 8(62), 105440-105457.
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8

Antibodies Used in Cell Signaling

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Antibodies used were HA (SC-805, Santa Cruz, Dallas, TX, USA), HA-7-HRP (H6533, Sigma-Aldrich), MEK1/2 (#9126, Cell Signaling, Danvers, MA, USA), phospho-MEK1/2 (#2338, Cell Signaling), ERK1/2 (M5670, Sigma-Aldrich), phospho- ERK1/2 (#4370, Cell Signaling), STAT5 (610191, BD Biosciences, Franklin Lakes, NJ, USA), phospho-STAT5A/B (05-886R, Merck Millipore), phospho-p70 S6 kinase (#9234, Cell Signaling), p70 S6 kinase (SC-230, Santa Cruz), RIPK3 (12107, Cell Signaling), HSP90 (610418, BD Transduction Laboratories), actin (AAN01-A, Cytoskeleton, Denver, CO, USA), and tubulin (ab7291, Abcam, Cambridge, UK). The secondary antibodies used were goat anti-mouse HRP (115-035-003, Jackson ImmunoResearch, West Grove, PA, USA), goat anti-rabbit HRP (111-035-003, Jackson ImmunoResearch), and Alexa Fluor 680 goat anti-mouse (A-21057, Molecular probes, Grand Island, NY, USA).
Bigenzahn J.W., Fauster A., Rebsamen M., Kandasamy R.K., Scorzoni S., Vladimer G.I., Müller A.C., Gstaiger M., Zuber J., Bennett K.L, & Superti-Furga G. (2015). An Inducible Retroviral Expression System for Tandem Affinity Purification Mass-Spectrometry-Based Proteomics Identifies Mixed Lineage Kinase Domain-like Protein (MLKL) as an Heat Shock Protein 90 (HSP90) Client. Molecular & Cellular Proteomics : MCP, 15(3), 1139-1150.
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9

Western Blot for Phospho-Protein Analysis

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Protein lysate was resolved by 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to a polyvinylidene difluoride membrane. The following Abs were used for immunoblotting: rabbit monoclonal Abs to phospho (p)-Akt (Ser473), Akt, extracellular signal-regulated kinases (Erk), p-Erk (Thr202/Tyr204) (all from Cell Signaling Technology, Danvers, MA, USA), p-STAT5 (EMD Millipore, Billerica, MA, USA), and STAT5 (BD Biosciences). The membranes were stained with appropriate Abs and visualized using SuperSignal West Femto Chemiluminescent Substrate (Thermo Fisher Scientific, Fremont, CA, USA).
Kim J.H., Sim J.H., Lee S., Seol M.A., Ye S.K., Shin H.M., Lee E.B., Lee Y.J., Choi Y.J., Yoo W.H., Kim J.H., Kim W.U., Lee D.S., Kim J.H., Kang I., Kang S.W, & Kim H.R. (2017). Interleukin-7 Induces Osteoclast Formation via STAT5, Independent of Receptor Activator of NF-kappaB Ligand. Frontiers in Immunology, 8, 1376.
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