- 44-382G - Invitrogen Antibodies STAT1 antibody

Submitted references Ruxolitinib inhibits cyclosporine-induced proliferation of cutaneous squamous cell carcinoma.

Abikhair Burgo M, Roudiani N, Chen J, Santana AL, Doudican N, Proby C, Felsen D, Carucci JA
JCI insight 2018 Sep 6;3(17)

Mediator Kinase Phosphorylation of STAT1 S727 Promotes Growth of Neoplasms With JAK-STAT Activation.

Nitulescu II, Meyer SC, Wen QJ, Crispino JD, Lemieux ME, Levine RL, Pelish HE, Shair MD
EBioMedicine 2017 Dec;26:112-125

RIPK1 ensures intestinal homeostasis by protecting the epithelium against apoptosis.

Takahashi N, Vereecke L, Bertrand MJ, Duprez L, Berger SB, Divert T, Gonçalves A, Sze M, Gilbert B, Kourula S, Goossens V, Lefebvre S, Günther C, Becker C, Bertin J, Gough PJ, Declercq W, van Loo G, Vandenabeele P
Nature 2014 Sep 4;513(7516):95-9

SUMO conjugation of STAT1 protects cells from hyperresponsiveness to IFNγ.

Begitt A, Droescher M, Knobeloch KP, Vinkemeier U
Blood 2011 Jul 28;118(4):1002-7

IL-4 and IL-13 negatively regulate TNF-alpha- and IFN-gamma-induced beta-defensin expression through STAT-6, suppressor of cytokine signaling (SOCS)-1, and SOCS-3.

Albanesi C, Fairchild HR, Madonna S, Scarponi C, De Pità O, Leung DY, Howell MD
Journal of immunology (Baltimore, Md. : 1950) 2007 Jul 15;179(2):984-92

The serine protease plasmin triggers expression of MCP-1 and CD40 in human primary monocytes via activation of p38 MAPK and janus kinase (JAK)/STAT signaling pathways.

Burysek L, Syrovets T, Simmet T
The Journal of biological chemistry 2002 Sep 6;277(36):33509-17

Protein tyrosine phosphatase 1B negatively regulates leptin signaling in a hypothalamic cell line.

Kaszubska W, Falls HD, Schaefer VG, Haasch D, Frost L, Hessler P, Kroeger PE, White DW, Jirousek MR, Trevillyan JM
Molecular and cellular endocrinology 2002 Sep 30;195(1-2):109-18

Protein tyrosine phosphatase 1B negatively regulates leptin signaling in a hypothalamic cell line.

Kaszubska W, Falls HD, Schaefer VG, Haasch D, Frost L, Hessler P, Kroeger PE, White DW, Jirousek MR, Trevillyan JM
Molecular and cellular endocrinology 2002 Sep 30;195(1-2):109-18

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Phospho-STAT1 (Ser727) using a polyclonal antibody (Product # 44-382G).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of STAT1 (pS727) 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 STAT1 (pS727) Rabbit polyclonal Antibody (Product # 44-382G) at 1:250 dilution in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (Product # A-11008) 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 DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (Product # A12381). Panel d is a merged image showing nuclear 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 STAT1 (pS727) 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 STAT1 (pS727) Rabbit Polyclonal Antibody (Product # 44-382G) 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 STAT1 (pS727) showing staining in the cytoplasm and nucleus of paraffin-embedded human cervical carcinoma (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 STAT1 (pS727) Rabbit Polyclonal Antibody (Product # 44-382G) 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: Flow cytometry analysis of STAT1 [pS727] was done on HeLa cells treated with IFN alpha (100ng/ml, 30 minutes). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with STAT1 [pS727] Rabbit Polyclonal Antibody (44382G, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor¨ 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ChIP- qPCR analysis of STAT1 (pSer727) was performed with 10 µL of the STAT1 (pSer727) Rabbit polyclonal antibody (Product # 44-382G) on sheared chromatin from 2 million HeLa cells treated with 50 ng/mL of IFN Gamma for one hour using the MAGnify Chromatin Immunoprecipitation System (Product # 49-2024). Normal Rabbit IgG 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 GLS1 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.

44-382G

Antibody data