Phospho-STAT3 (Ser727) Monoclonal antibody

Phospho-STAT3 (Ser727) Monoclonal Antibody for WB, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG2a

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

2G11G11

Cat no : 60479-1-Ig

Synonyms

p STAT3 (Ser727), p STAT3, p STAT3 Ser727, Phospho STAT3, Phospho STAT3 (Ser727)

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Validation Data Gallery

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  WB analysis using 60479-1-Ig

  UV treated and untreated HeLa cells were subjected to SDS PAGE followed by western blot with 60479-1-Ig (Phospho-STAT3 (Ser727) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Alpha Tubulin (HRP-66031) antibody as a loading control.

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  WB analysis using 60479-1-Ig

  EGF treated and untreated A549 cells were subjected to SDS PAGE followed by western blot with 60479-1-Ig (Phospho-STAT3 (Ser727) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Alpha Tubulin (HRP-66031) antibody as a loading control.

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  WB analysis using 60479-1-Ig

  TNF alpha (HZ-1014) treated and untreated MCF-7 cells were subjected to SDS PAGE followed by western blot with 60479-1-Ig (Phospho-STAT3 (Ser727) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Alpha Tubulin (HRP-66031) antibody as a loading control.

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  WB analysis using 60479-1-Ig

  TNF alpha (HZ-1014) treated and untreated NIH/3T3 cells were subjected to SDS PAGE followed by western blot with 60479-1-Ig (Phospho-STAT3 (Ser727) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with HRP-conjugated Alpha Tubulin (HRP-66031) antibody as a loading control.

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  WB analysis of HSC-T6 using 60479-1-Ig

  UV treated HSC-T6 cells were subjected to SDS PAGE followed by western blot with 60479-1-Ig (STAT3 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IF Staining of HeLa using 60479-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HeLa cells using Phospho-STAT3 (Ser727) antibody (60479-1-Ig, Clone: 2G11G11 ) at dilution of 1:500 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

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  IF Staining of HeLa using 60479-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed untreated HeLa cells, UV (1 hour) and 100 nM Calyculin A (30 minutes) treated HeLa cells using Phospho-STAT3 (Ser727) antibody (60479-1-Ig, Clone: 2G11G11) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

• 

  FC experiment of HeLa using 60479-1-Ig

  1x10^6 untreated HeLa cells (dash lines), UV (1 hour) and 100 nM Calyculin A (30 minutes) treated HeLa cells (full lines) were intracellularly stained with 0.1 μg Phospho-STAT3 (Ser727) Monoclonal antibody (60479-1-Ig, Clone: 2G11G11, red) and Multi-rAb CoraLite ® Plus 647-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM005). Mouse IgG2a isotype control Mouse McAb (66360-2-Ig, Clone: 11A1B2, blue) was parallel stained as control. Cells were fixed with 4% PFA .

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Tested Applications

• Positive WB detected in: HeLa cells, A549 cells, UV treated HSC-T6 cells, MCF-7 cells, NIH/3T3 cells, EGF treated A549 cells, UV treated HeLa cells, TNF alpha treated MCF-7 cells
• Positive IF/ICC detected in: UV(1hour),100 nM Calyculin A(30 minutes)treated HeLa cells, HeLa cells
• Positive FC (Intra) detected in: UV (1 hour) and 100 nM Calyculin A (30 minutes) HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:500-1:2000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.10 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

60479-1-Ig targets Phospho-STAT3 (Ser727) in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Mouse / IgG2a
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: signal transducer and activator of transcription 3 (acute-phase response factor)
• Calculated Molecular Weight: 770 aa, 88 kDa
• Observed Molecular Weight: 85-90 kDa
• GenBank Accession Number: BC000627
• Gene Symbol: STAT3
• Gene ID (NCBI): 6774
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Signal transducer and activator of transcription 3 (acute-phase response factor) (STAT3, synonyms: APRF, FLJ20882, MGC16063) is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. STAT3 is activated through phosphorylation in response to various cytokines and growth factors including IFNs, EGF, IL5, IL6, HGF, LIF and BMP2. STAT3 mediates the expression of a variety of genes in response to cell stimuli, and thus plays a key role in many cellular processes such as cell growth and apoptosis.