- 13-5300 - Invitrogen Antibodies STAT5B antibody

Submitted references IFN-β rescues neurodegeneration by regulating mitochondrial fission via STAT5, PGAM5, and Drp1.

Tresse E, Riera-Ponsati L, Jaberi E, Sew WQG, Ruscher K, Issazadeh-Navikas S
The EMBO journal 2021 Jun 1;40(11):e106868

Inhibition of the JAK/STAT Signaling Pathway in Regulatory T Cells Reveals a Very Dynamic Regulation of Foxp3 Expression.

Goldstein JD, Burlion A, Zaragoza B, Sendeyo K, Polansky JK, Huehn J, Piaggio E, Salomon BL, Marodon G
PloS one 2016;11(4):e0153682

DNA methylation and transcription in a distal region upstream from the bovine AlphaS1 casein gene after once or twice daily milking.

Nguyen M, Boutinaud M, Pétridou B, Gabory A, Pannetier M, Chat S, Bouet S, Jouneau L, Jaffrezic F, Laloë D, Klopp C, Brun N, Kress C, Jammes H, Charlier M, Devinoy E
PloS one 2014;9(11):e111556

Identification of STAT5A and STAT5B target genes in human T cells.

Kanai T, Seki S, Jenks JA, Kohli A, Kawli T, Martin DP, Snyder M, Bacchetta R, Nadeau KC
PloS one 2014;9(1):e86790

Prolactin signalling in the mouse hypothalamus is primarily mediated by signal transducer and activator of transcription factor 5b but not 5a.

Yip SH, Eguchi R, Grattan DR, Bunn SJ
Journal of neuroendocrinology 2012 Dec;24(12):1484-91

Identification of human STAT5-dependent gene regulatory elements based on interspecies homology.

Nelson EA, Walker SR, Li W, Liu XS, Frank DA
The Journal of biological chemistry 2006 Sep 8;281(36):26216-24

Activated signal transducer and activator of transcription (STAT) 3: localization in focal adhesions and function in ovarian cancer cell motility.

Silver DL, Naora H, Liu J, Cheng W, Montell DJ
Cancer research 2004 May 15;64(10):3550-8

Stat5a is tyrosine phosphorylated and nuclear localized in a high proportion of human breast cancers.

Cotarla I, Ren S, Zhang Y, Gehan E, Singh B, Furth PA
International journal of cancer 2004 Feb 20;108(5):665-71

Isolation of unique STAT5 targets by chromatin immunoprecipitation-based gene identification.

Nelson EA, Walker SR, Alvarez JV, Frank DA
The Journal of biological chemistry 2004 Dec 24;279(52):54724-30

Hormone-induced modifications of the chromatin structure surrounding upstream regulatory regions conserved between the mouse and rabbit whey acidic protein genes.

Millot B, Montoliu L, Fontaine ML, Mata T, Devinoy E
The Biochemical journal 2003 May 15;372(Pt 1):41-52

Suppressor of cytokine signaling-1 is a critical regulator of interleukin-7-dependent CD8+ T cell differentiation.

Chong MM, Cornish AL, Darwiche R, Stanley EG, Purton JF, Godfrey DI, Hilton DJ, Starr R, Alexander WS, Kay TW
Immunity 2003 Apr;18(4):475-87

Suppressor of cytokine signaling-1 is a critical regulator of interleukin-7-dependent CD8+ T cell differentiation.

Chong MM, Cornish AL, Darwiche R, Stanley EG, Purton JF, Godfrey DI, Hilton DJ, Starr R, Alexander WS, Kay TW
Immunity 2003 Apr;18(4):475-87

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of endogenous STAT5a and STAT5b expression in HeLa cells using Rb x STAT5a and Rb x STAT5b.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockout of STAT5B was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR773180_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of STAT5B was performed by loading 30 µg of HeLa wild type (Lane 1) and HeLa STAT5B KO (Lane 2) modified whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-STAT5 beta Monoclonal Antibody (ST5b-10G1) (Product # 13-5300, 1:250 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to STAT5B.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of STAT5 beta was performed by loading 20 µg of NIH/3T3 (lane1), HeLa (lane2), K562 (lane3), Ramos (lane4) and Jurkat (lane5) cell lysate using Novex® NuPAGE® 12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (LC5800), and iBlot® Dry Blotting System (IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5 % skim milk for 1 hour at room temperature. STAT5 beta was detected at ~ 92 kDa using STAT5 beta Mouse Monoclonal Antibody (Product # 13-5300) at 1:250 dilution in 2.5 % skim milk at 4°C overnight on a rocking platform. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1:5000 dilution was used and chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of STAT5 beta was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with STAT5 beta Mouse Monoclonal Antibody (Product # 13-5300) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Rabbit Anti-Mouse IgG Secondary Antibody (Product # A-11059) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of STAT5 beta showing staining in the cytoplasm and nucleus of paraffin-embedded human T cell lymphoma (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a STAT5 beta monoclonal antibody (Product # 13-5300) diluted in 3% BSA-PBS at a dilution of 1:100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of STAT5 beta showing staining in the cytoplasm and nucleus of paraffin-embedded mouse lymph node (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a STAT5 beta monoclonal antibody (Product # 13-5300) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of STAT5 beta showing staining in the cytoplasm and nucleus of paraffin-embedded mouse spleen tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a STAT5 beta monoclonal antibody (Product # 13-5300) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ChIP- qPCR analysis of STAT5 beta was performed with 5 µg/mL of the STAT5 beta Mouse monoclonal antibody (Product # 13-5300) on sheared chromatin from 2 million HeLa-IFN Gamma treated (1hr) cells using the MAGnify™ Chromatin Immunoprecipitation System (Product # 49-2024).Normal mouse IgG (5 µg/mL) was used as a negative IP control. The purified DNA from each ChIP sample was analyzed by StepOnePlus™ Real-Time PCR System (Product # 4376600) with primers for the promoter of active AP1, COX1 gene, used as positive control target, and the inactive SAT2, used as negative control target. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 Binding activity of two potential STAT5 binding sites located within the CSN1S1 distal upstream region. (A) Representative EMSA. Mammary nuclear extract (22ug) from rabbits in early lactation were incubated with the indicated labelled 32 P probes in the presence or absence of STAT5a or STAT5b antibodies (1 or 0.5 ug, respectively). Complexes were analysed on non denaturating 5% acrylamide gels in 0.25xTBE. The lower panel is an overexposed image of the upper one to show the weak interaction between D1 and nuclear extracts and the supershifts with STAT5 antibodies. (B) Competition assays with increasing amounts of unlabelled C, D2, D1 and D1m using 32 P-C or 32 P-D2, are indicated in each graph. Results are expressed as a percentage of the radioactivity bound in the absence of unlabelled oligonucleotides and plotted as a function of the log (ng oligonucleotide) in the reaction.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: EV5 Figure Impact of modulation in STATs and PGAM5 on IFN-beta-dependent Drp1 phosphorylation and mitochondrial health Immunoblot of total and phospho-S622-Drp1 in wild-type N2A cells or N2A cells silenced for STAT-2 or CDK5. Vinculin was used as a loading control. CDK5 was used as a positive control for the loss of Drp1 phosphorylation. Immunoblot of total or phospho-S622-Drp1 in wild-type CNs or CNs silenced for STAT1, STAT2, or STAT3. beta3-tubulin was used as a loading control. STAT5B immunostaining in N2A cells depleted for STAT5 A and B by CRISPR/Cas9 ( deltaStat5 ) or with nontargeting control (NTC). PGAM5, CamKIIa, ERK2, and Cdk5 expression in Ifnb +/+ and Ifnb -/- CGNs extracted from microarrays. Immunoblot for STAT5, pan- and phospho-Drp1, and PGAM5 on rIFN-beta, IL-2, and IL-3 treatment in wild-type CNs at DIV6. Quantification of (E). Immunoblot for PGAM5 and quantification in Ifnb +/+ and Ifnb -/- CNs at DIV6. immunostaining for PGAM5 and beta3-tubulin in Ifnb +/+ and Ifnb -/- CNs with or without rIFN-beta. Nuclei stained with DAPI. Immunoblot in N2A control or N2A cells transfected with si-NTC or si-ERK2 with or without rIFN-beta for 2 h. Immunoblot for pan- and phosphor-Drp1 and PGAM5 in N2A cells transfected with si-CamKIIa with rIFN-beta for 0 to 4 h. Matching si-NTC control is presented in Fig 6, and side-by-side blots with anti-CamKIIa pan and phosphorylated from the same untreated samples are provided in the upper panel. Immunoblot for STAT5 and pan- and ph

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1 Localization of STAT5A and STAT5B, and monomers and dimers of STAT5A and STAT5B. A demonstrates translocation of STAT5A and STAT5B into cell nuclei after 30-2 (40x confocal). Yellow, STAT5A; purple, STAT5B; blue, nucleus. B and C . Detection of STAT5A and STAT5B proteins in cytoplasmic or nuclear proteins fractionated from CD4 + T cells after PHA-P stimulation for 3 days followed by incubation with rhIL-2 for 30 min. B . STAT5A monomer (91 kDa, arrow 1) and STAT5A dimer (arrow 2) in native cytoplasmic or nuclear proteins, detected with anti-STAT5A Ab. C . STAT5B monomer (90 kDa, arrow 3) and STAT5B dimer (arrow 4) in native cytoplasmic or nuclear proteins, detected using anti-STAT5B Ab. D . Detection of phosphorylated STAT5 proteins in STAT5A- or STAT5B- immunoprecipitated nuclear proteins fractionated from CD4 + T cells after PHA-P stimulation for 3 days followed by incubation with rh-IL-2 for 0 min, 30 min or 3 days. E . Detection of phosphorylated STAT5 proteins in cytoplasmic or nuclear proteins fractionated from CD4 + T cells after PHA-P stimulation for 3 days followed by incubation with rhIL-2 for 3 days, and control (unstimulated condition).

13-5300

Antibody data