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STAT1 alpha Recombinant Rabbit Monoclonal Antibody [SJ01-89]

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Catalog# ET1606-39

STAT1 alpha Recombinant Rabbit Monoclonal Antibody [SJ01-89]

Application

  • WB

  • IP

  • FC

  • IHC-P

  • IF-Cell

  • IF-Tissue

Reactivity

  • Human

  • Mouse

This product has been cited in peer reviewed publications, see list HERE

概述

产品名称

STAT1 alpha Recombinant Rabbit Monoclonal Antibody [SJ01-89]

抗体类型

Recombinant Rabbit monoclonal Antibody

免疫原

Synthetic peptide within human STAT1 aa 710-750.

种属反应性

Human, Mouse

验证应用

WB, IP, FC, IHC-P, IF-Cell, IF-Tissue

分子量

Predicted band size: 87 kDa

阳性对照

Jurkat cell lysate, A431 cell lysate, HeLa cell lysate, A549 cell lysate, SK-Br-3 cell lysate, SK-MEL-28 cell lysate, HT-29 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, human colon tissue, human ovary cancer tissue, human spleen tissue, mouse colon tissue, MCF-7.

偶联

unconjugated

克隆号

SJ01-89

RRID

AB_3069734

产品特性

形态

Liquid

浓度

1ug/ul

存放说明

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

存储缓冲液

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

亚型

IgG

纯化方式

Protein A affinity purified.

应用稀释度

  • WB

  • 1:1,000-1:5,000

  • FC

  • 1:50-1:100

  • IP

  • Use at an assay dependent concentration.

  • IHC-P

  • 1:500-1:2,000

  • IF-Cell

  • 1:100

  • IF-Tissue

  • 1:50-1:400

发表文章中的应用

WB See 9 publications below
CO-IP See 1 publications below

发表文章中的种属

Mouse See 6 publications below
Human See 5 publications below

靶点

功能

Membrane receptor signaling by various ligands, including interferons and growth hormones such as EGF, induces activation of JAK kinases which then leads to tyrosine phosphorylation of the various Stat transcription factors. Stat1 and Stat2 are induced by IFN-α and form a heterodimer which is part of the ISGF3 transcription factor complex. Although early reports indicate Stat3 activation by EGF and IL-6, it has been shown that Stat3β appears to be activated by both while Stat3α is activated by EGF, but not by IL-6. Highest expression of Stat4 is seen in testis and myeloid cells. IL-12 has been identified as an activator of Stat4. Stat5 has been shown to be activated by Prolactin and by IL-3. Stat6 is involved in IL-4 activated signaling pathways. Mutations in the STAT1 molecule can be gain of function (GOF) or loss of function (LOF). Both of them can cause different phenotypes and symptoms. Recurring common infections are frequent in both GOF and LOF mutations. In humans STAT1 has been particularly under strong purifying selection when populations shifted from hunting and gathering to farming, because this went along with a change in the pathogen spectrum. STAT1 loss of function, therefore STAT1 deficiency can have many variants. There are two main genetic impairments that can cause response to interferons type I and III. First there can be autosomal recessive partial or even complete deficiency of STAT1. That causes intracellular bacterial diseases or viral infections and impaired IFN a, b, g and IL27 responses are diagnosed. In partial form there can also be found high levels of IFNg in blood serum. When tested from whole blood, monocytes do not respond to BCG and IFNg doses with IL-12 production. In complete recessive form there is a very low response to anti-viral and antimycotical medication. Second, partial STAT1 deficiency can also be an autosomal dominant mutation; phenotypically causing impaired IFNg responses and causing patients to suffer with selective intracellular bacterial diseases (MSMD).

背景文献

1. Shakya R et al. Hypomethylating therapy in an aggressive stroma-rich model of pancreatic carcinoma. Cancer Res 73:885-96 (2013).

2. Syu LJ et al. Transgenic expression of interferon- in mouse stomach leads to inflammation, metaplasia, and dysplasia. Am J Pathol 181:2114-25 (2012).

序列相似性

Belongs to the transcription factor STAT family.

翻译后修饰

Phosphorylated on tyrosine and serine residues in response to a variety of cytokines/growth hormones including IFN-alpha, IFN-gamma, PDGF and EGF. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Upon EGF stimulation, phosphorylation on Tyr-701 (lacking in beta form) by JAK1, JAK2 or TYK2 promotes dimerization and subsequent translocation to the nucleus. Growth hormone (GH) activates STAT1 signaling only via JAK2. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4. Phosphorylation on Ser-727 by several kinases including MAPK14, ERK1/2 and CAMKII on IFN-gamma stimulation, regulates STAT1 transcriptional activity. Phosphorylation on Ser-727 promotes sumoylation though increasing interaction with PIAS. Phosphorylation on Ser-727 by PRKCD induces apoptosis in response to DNA-damaging agents. Phosphorylated on tyrosine residues when PTK2/FAK1 is activated; most likely this is catalyzed by a SRC family kinase. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates interferon-mediated signaling. Upon viral infection or IFN induction, phosphorylation on Ser-708 occurs much later than phosphorylation on Tyr-701 and is required for the binding of ISGF3 on the ISREs of a subset of IFN-stimulated genes IKBKE-dependent. Phosphorylation at Tyr-701 and Ser-708 are mutually exclusive, phosphorylation at Ser-708 requires previous dephosphorylation of Tyr-701.; Sumoylated with SUMO1, SUMO2 and SUMO3. Sumoylation is enhanced by IFN-gamma-induced phosphorylation on Ser-727, and by interaction with PIAS proteins. Enhances the transactivation activity.; ISGylated.; Mono-ADP-ribosylated at Glu-657 and Glu-705 by PARP14; ADP-ribosylation prevents phosphorylation at Tyr-701. However, the role of ADP-ribosylation in the prevention of phosphorylation has been called into question and the lack of phosphorylation may be due to sumoylation of Lys-703.; Monomethylated at Lys-525 by SETD2; monomethylation is necessary for phosphorylation at Tyr-701, translocation into the nucleus and activation of the antiviral defense.

亚细胞定位

Cytoplasm, Nucleus.

UNIPROT #

SwissProt: P42224 Human

SwissProt: P42225 Mouse

别名

Signal transducer and activator of transcription 1 91kD antibody

CANDF7 antibody

DKFZp686B04100 antibody

IMD31A antibody

IMD31B antibody

IMD31C antibody

ISGF 3 antibody

ISGF-3 antibody

OTTHUMP00000163552 antibody

OTTHUMP00000165046 antibody

展开

图片

  • Western blot analysis of STAT1 alpha on different lysates with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/1,000 dilution.<br /><br />Lane 1: Jurkat cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: HeLa cell lysate<br />Lane 4: A549 cell lysate<br />Lane 5: SK-Br-3 cell lysate<br />Lane 6: SK-MEL-28 cell lysate<br />Lane 7: HT-29 cell lysate<br />Lane 8: RAW264.7 cell lysate<br />Lane 9: C2C12 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 87 kDa<br />Observed band size: 87 kDa<br /><br />Exposure time: 14 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of STAT1 alpha on different lysates with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/1,000 dilution.

    Lane 1: Jurkat cell lysate
    Lane 2: A431 cell lysate
    Lane 3: HeLa cell lysate
    Lane 4: A549 cell lysate
    Lane 5: SK-Br-3 cell lysate
    Lane 6: SK-MEL-28 cell lysate
    Lane 7: HT-29 cell lysate
    Lane 8: RAW264.7 cell lysate
    Lane 9: C2C12 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 87 kDa
    Observed band size: 87 kDa

    Exposure time: 14 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-39) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of STAT1 alpha on different lysates with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/5,000 dilution.<br /><br />Lane 1: HAP1-parental cell lysate<br />Lane 2: HAP1-STAT1 alpha KD cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 87 kDa<br />Observed band size: 87 kDa<br /><br />Exposure time: 60 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/5,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of STAT1 alpha on different lysates with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/5,000 dilution.

    Lane 1: HAP1-parental cell lysate
    Lane 2: HAP1-STAT1 alpha KD cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 87 kDa
    Observed band size: 87 kDa

    Exposure time: 60 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1606-39) at 1/5,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-39) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-39) at 1/500 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-39) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/2,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/2,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1606-39) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Flow cytometric analysis of STAT1 alpha was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of STAT1 alpha was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1606-39, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Immunocytochemistry analysis of HeLa cells labeling STAT1 alpha with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling STAT1 alpha with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of RAW264.7 cells labeling STAT1 alpha with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-STAT1 alpha antibody (<a href="/products/ET1606-39" style="font-weight: bold;text-decoration: underline;">ET1606-39</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of RAW264.7 cells labeling STAT1 alpha with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-STAT1 alpha antibody (ET1606-39) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

引文

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