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Phospho-STAT3 (Y705) Recombinant Rabbit Monoclonal Antibody [SZ43-01]

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Specification

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Catalog# ET1603-40

Phospho-STAT3 (Y705) Recombinant Rabbit Monoclonal Antibody [SZ43-01]

Application

  • WB

  • IHC-P

  • IP

Reactivity

  • Human

  • Mouse

BSA and Azide free
Predicted reactivity

  • Rat

Predicted species support after-sales service
Conjugation

  • unconjugated

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

Overview

Product Name

Phospho-STAT3 (Y705) Recombinant Rabbit Monoclonal Antibody [SZ43-01]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Tyr705 of human STAT3.

Product Specificity

ET1603-40 Rabbit mAb detects phosphorylation of STAT3 at Y705, as well as phosphorylation of the STAT3_ΔS701 isoform at the same residue.

Species Reactivity

Human, Mouse (Predicted: Rat)

Predicted species support after-sales service

Validated Applications

WB, IHC-P, IP

Molecular Weight

Predicted band size: 88 kDa

Positive Control

HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate, Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate, A431 treated with 100ng/mL EGF for 30 minutes cell lysate, human lung tissue, M‑NFS‑60 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, L6 cell lysate, Mouse heart tissue lysate, SK-Br-3 cell lysates, human kidney tissue.

Conjugation

unconjugated

Clone Number

SZ43-01

RRID

AB_3069682

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

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

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IHC-P

  • 1:500

  • IP

  • 1-2μg/sample

Target

Function

Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene. May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Acts as a regulator of inflammatory response by regulating differentiation of naive CD4+ T-cells into T-helper Th17 or regulatory T-cells (Treg): deacetylation and oxidation of lysine residues by LOXL3, leads to disrupt STAT3 dimerization and inhibit its transcription activity. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1. Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation. May play an apoptotic role by transctivating BIRC5 expression under LEP activation. Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. Plays a crucial role in basal beta cell functions, such as regulation of insulin secretion.

Background References

1. Andersson KM et al. Down-regulation of survivin alleviates experimental arthritis. J Leukoc Biol 97:135-45 (2015).

2. Schwartz C et al. Melatonin receptor signaling contributes to neuroprotection upon arousal from torpor in thirteen-lined ground squirrels. Am J Physiol Regul Integr Comp Physiol 309:R1292-300 (2015).

Sequence Similarity

Belongs to the transcription factor STAT family.

Tissue Specificity

Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas.

Post-translational Modification

Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling.; Acetylated on lysine residues by CREBBP. Deacetylation by LOXL3 leads to disrupt STAT3 dimerization and inhibit STAT3 transcription activity. Oxidation of lysine residues to allysine on STAT3 preferentially takes place on lysine residues that are acetylated.; Some lysine residues are oxidized to allysine by LOXL3, leading to disrupt STAT3 dimerization and inhibit STAT3 transcription activity. Oxidation of lysine residues to allysine on STAT3 preferentially takes place on lysine residues that are acetylated.; (Microbial infection) Phosphorylated on Tyr-705 in the presence of S.typhimurium SarA.; S-palmitoylated by ZDHHC19 in SH2 putative lipid-binding pockets, leading to homodimerization. Nuclear STAT3 is highly palmitoylated (about 75%) compared with cytoplasmic STAT3 (about 20%).; S-stearoylated, probably by ZDHHC19.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P40763 Human

SwissProt: P42227 Mouse

SwissProt: P52631 Rat

Synonyms

1110034C02Rik antibody

Acute Phase Response Factor antibody

Acute-phase response factor antibody

ADMIO antibody

APRF antibody

AW109958 antibody

DNA binding protein APRF antibody

FLJ20882 antibody

HIES antibody

MGC16063 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (<a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate<br />Lane 3: Jurkat cell lysate<br />Lane 4: Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate<br />Lane 5: A431 cell lysate<br />Lane 6: A431 treated with 100ng/mL EGF for 30 minutes cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 88 kDa<br />Observed band size: 88 kDa<br /><br />Exposure time: 35 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/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate
    Lane 3: Jurkat cell lysate
    Lane 4: Jurkat treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate
    Lane 5: A431 cell lysate
    Lane 6: A431 treated with 100ng/mL EGF for 30 minutes cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 88 kDa
    Observed band size: 88 kDa

    Exposure time: 35 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 (ET1603-40) at 1/5,000 dilution and competitor's antibody at 1/1,000 dilution were 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.

  • Immunohistochemical analysis of paraffin-embedded human lung tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (<a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</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 lung tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1603-40) 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.

  • Western blot analysis of Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (<a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/1,000 dilution.<br /><br />Lane 1: M‑NFS‑60 cell lysate (20 µg/Lane)<br />Lane 2: RAW264.7 cell lysate (20 µg/Lane)<br />Lane 3: C2C12 cell lysate (20 µg/Lane)<br />Lane 4: L6 cell lysate (20 µg/Lane)<br />Lane 5: Mouse heart tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 88 kDa<br />Observed band size: 88 kDa<br /><br />Exposure time: 25 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/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</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 Phospho-STAT3 (Y705) on different lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/1,000 dilution.

    Lane 1: M‑NFS‑60 cell lysate (20 µg/Lane)
    Lane 2: RAW264.7 cell lysate (20 µg/Lane)
    Lane 3: C2C12 cell lysate (20 µg/Lane)
    Lane 4: L6 cell lysate (20 µg/Lane)
    Lane 5: Mouse heart tissue lysate (40 µg/Lane)

    Predicted band size: 88 kDa
    Observed band size: 88 kDa

    Exposure time: 25 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 (ET1603-40) 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.

  • Western blot analysis of Phospho-STAT3 (Y705) on SK-Br-3 cell lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (<a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/500 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 88 kDa<br />Observed band size: 88 kDa<br /><br />Exposure time: 2 minutes;<br /><br />8% 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/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:300,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-STAT3 (Y705) on SK-Br-3 cell lysates with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution.

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 88 kDa
    Observed band size: 88 kDa

    Exposure time: 2 minutes;

    8% 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 (ET1603-40) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (<a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a>) at 1/500 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</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 kidney tissue with Rabbit anti-Phospho-STAT3 (Y705) antibody (ET1603-40) at 1/500 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (ET1603-40) 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.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Phospho-STAT3 (Y705) was immunoprecipitated from 0.2 mg HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate with <a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate (input)<br />Lane 2: Rabbit IgG instead of <a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a> in HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate<br />Lane 3: <a href="/products/ET1603-40" style="font-weight: bold;text-decoration: underline;">ET1603-40</a> IP in HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 24 seconds; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    ☑ Cell treatment (CT)

    Phospho-STAT3 (Y705) was immunoprecipitated from 0.2 mg HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate with ET1603-40 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1603-40 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate (input)
    Lane 2: Rabbit IgG instead of ET1603-40 in HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate
    Lane 3: ET1603-40 IP in HeLa treated with 50ng/mL IFN alpha 1 for 30 minutes cell lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 24 seconds; ECL: K1802

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

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