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Signal Transducer And Activator Of Transcription 3 Peptide 2

Antibody: CPTC-STAT3-2
Background

Catalog Number:

CPTC-STAT3-2

Target Antigen:

Signal Transducer And Activator Of Transcription 3 Peptide 2

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

04/09/2024

Antigen Recognition(s):

Peptide, Recombinant Full-length, Endogenous, Phosphorylation

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Characterization Data [Compare Characterization Data]
  Affinity Measurement by Biolayer Interferometry
Click to enlarge image The affinity and binding kinetics of CPTC-STAT3-2 antibody and human signal transducer and activator of transcription 3 (acute-phase response factor) (STAT3) recombinant protein, transcript variant 1, were measured using biolayer interferometry. Recombinant protein was covalently immobilized onto AR2G biosensors using standard amine coupling. CPTC-STAT3-2 antibody at 1024 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without recombinant protein were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent model. Click image to enlarge

CPTC-STAT3-2 Affinity and Kinetics (Bio-Layer Interferometry)

Result: Positive

The affinity and binding kinetics of CPTC-STAT3-2 antibody and human signal transducer and activator of transcription 3 (acute-phase response factor) (STAT3) recombinant protein, transcript variant 1, were measured using biolayer interferometry. Recombinant protein was covalently immobilized onto AR2G biosensors using standard amine coupling. CPTC-STAT3-2 antibody at 1024 nM, 64 nM, 16 nM, 4 nM, 1 nM, and 0.25 nM, was used as analyte. Buffer only and biosensors immobilized without recombinant protein were used as references for background subtraction. All data was analyzed globally using a 1:2 bivalent model.


Characterization SOP Files

  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-STAT3-2 and human signal transducer and activator of transcription 3 (acute-phase response factor) (STAT3) recombinant protein, transcript variant 1, were measured using surface plasmon resonance. Recombinant protein was amine coupled onto a series S CM5 biosensor chip. CPTC-STAT3-2 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent model. Click image to enlarge

CPTC-STAT3-2 Affinity and Kinetics (Surface Plasmon Resonance) 

Result: Positive

Affinity and binding kinetics of CPTC-STAT3-2 and human signal transducer and activator of transcription 3 (acute-phase response factor) (STAT3) recombinant protein, transcript variant 1, were measured using surface plasmon resonance. Recombinant protein was amine coupled onto a series S CM5 biosensor chip. CPTC-STAT3-2 antibody at 1024 nM, 256 nM, 64 nM, 16 nM, 4 nM, 1.0 nM and 0.25 nM, was used as analyte. Binding data were double-referenced and analyzed globally using a bivalent model.


Characterization SOP Files

  Flow Cytometry
Click to enlarge image Flow cytometric analysis of STAT3 expression in SN12C and COLO205 cells using CPTC-STAT3-2 rabbit antibody. SN12C cells were permeabilized, fixed and then stained with CPTC-STAT3-2 antibody (solid green) or concentration-matched rabbit isotype control antibody (dashed green). COLO205 cells were permeabilized, fixed and then stained with CPTC-STAT3-2 (solid blue) or concentration-matched rabbit isotype control antibody (dashed blue) and then fixed. A BV421 conjugated goat anti-rabbit IgG (H+L) was used as a secondary antibody. All data were analyzed using FlowJo.  CPTC-STAT3-2 antibody can detect expression of STAT3 in both SN12C and COLO205 cells. Click image to enlarge

CPTC-STAT3-2 (Flow Cytometry)

Result: Positive

Flow cytometric analysis of STAT3 expression in SN12C and COLO205 cells using CPTC-STAT3-2 rabbit antibody. SN12C cells were permeabilized, fixed and then stained with CPTC-STAT3-2 antibody (solid green) or concentration-matched rabbit isotype control antibody (dashed green). COLO205 cells were permeabilized, fixed and then stained with CPTC-STAT3-2 (solid blue) or concentration-matched rabbit isotype control antibody (dashed blue) and then fixed. A BV421 conjugated goat anti-rabbit IgG (H+L) was used as a secondary antibody. All data were analyzed using FlowJo.  CPTC-STAT3-2 antibody can detect expression of STAT3 in both SN12C and COLO205 cells.


Characterization SOP Files

  IHC Tissue
Click to enlarge image Tissue Micro-Array (TMA) core of prostate cancer showing cytoplasmic and membranous staining using Antibody CPTC-STAT3-2. Titer: 1:100 Click image to enlarge

CPTC-STAT3-2 IHC Tissue

Result: Positive

Tissue Micro-Array (TMA) core of prostate cancer showing cytoplasmic and membranous staining using Antibody CPTC-STAT3-2. Titer: 1:100


Characterization SOP Files

  IP
Click to enlarge image Immunoprecipitation using CPTC-STAT3-2 as capture antibody to probe the lysates of SNC12, IGROv1 and MCF7. Eluates were tested in Simple Western, using CPTC-STA3-2 as primary antibody (Panel A). The antibosy was not able to pull down the target protein, as also evident in the comparison between input material (blue line) and eluate (green line) profiles in panels B, C and D, respectivelly for SNC12C, IGROV1 and MCF7. Click image to enlarge

CPTC-STA3-2 IP_SW (Lysates)

Result: Negative

Immunoprecipitation using CPTC-STAT3-2 as capture antibody to probe the lysates of SNC12, IGROv1 and MCF7. Eluates were tested in Simple Western, using CPTC-STA3-2 as primary antibody (Panel A). The antibosy was not able to pull down the target protein, as also evident in the comparison between input material (blue line) and eluate (green line) profiles in panels B, C and D, respectivelly for SNC12C, IGROV1 and MCF7.


Characterization SOP Files

  Immunofluorescence

CPTC-STAT3-2 Immunofluorescence

Result: Negative

Immunofluorescence staining of human cell line SN12C using CPTC-STAT3-2 as primary antibody. STAT3 protein expression shows no localization.


Characterization SOP Files

  NCI 60 Protein Array

CPTC-STAT3-2 NCI 60 Protein Array

Result: Negative

This antibody is not suitable for use in a Reverse Phase Protein Array format as described in SOP M-105.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Western Blot using CPTC-STAT3-2 as primary antibody against the over-expressed lysate of STAT3 (expected MW is 88 KDa). The antibody is able to recognize the target protein. Click image to enlarge

CPTC-STAT3-2 Western Blot (over-expressed lysate)

Result: Positive

Western Blot using CPTC-STAT3-2 as primary antibody against the over-expressed lysate of STAT3 (expected MW is 88 KDa). The antibody is able to recognize the target protein.


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Simple Western (automated WB) using CPTC-STAT3-2 as primary antibody against the over-expressed lysate of STAT3 (expected MW is 88 KDa). The protein in the over-expressed lysate is Myc-tagged, and it was also tested with a anti-Myc antibody (positive control, bottom panel). The antibody CPTC-STAT3-3 is able to recognize the target protein in the over-expressed lysate. Click image to enlarge

CPTC-STAT3-2 Simple Western (over-expressed lysate)

Result: Positive

Simple Western (automated WB) using CPTC-STAT3-2 as primary antibody against the over-expressed lysate of STAT3 (expected MW is 88 KDa). The protein in the over-expressed lysate is Myc-tagged, and it was also tested with a anti-Myc antibody (positive control, bottom panel). The antibody CPTC-STAT3-3 is able to recognize the target protein in the over-expressed lysate.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western Blot using CPTC-STAT3-2 as primary antibody against cell lysate of SN12C, COLO205, IGROV1, NCI H460 and MCF7. Expected MW is 88 KDa. The antibody is able to recognize the target protein in the whole lysates of SN12C, IGROV1, and weakly in MCF7. Click image to enlarge

CPTC-STAT3-2 Western Blot (cell lysates)

Result: Positive

Western Blot using CPTC-STAT3-2 as primary antibody against cell lysate of SN12C, COLO205, IGROV1, NCI H460 and MCF7. Expected MW is 88 KDa. The antibody is able to recognize the target protein in the whole lysates of SN12C, IGROV1, and weakly in MCF7.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Simple Western (automated WB) using CPTC-STAT3-2 as primary antibody against the cell lysate of SN12C, COLO205, IGROV1, NCI H460 and MCF7. Expected MW is 88 KDa. The antibody is able to recognize the target protein in the whole lysates of SN12C, IGROV1, and weakly in MCF7. The quality of the lysates was verified probing them with an anti-vinculin antibody (loading control, bottom panel). Click image to enlarge

CPTC-STAT3-2 Simple Western (cell lysate)

Result: Positive

Simple Western (automated WB) using CPTC-STAT3-2 as primary antibody against the cell lysate of SN12C, COLO205, IGROV1, NCI H460 and MCF7. Expected MW is 88 KDa. The antibody is able to recognize the target protein in the whole lysates of SN12C, IGROV1, and weakly in MCF7. The quality of the lysates was verified probing them with an anti-vinculin antibody (loading control, bottom panel).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Single cell western blot using CPTC-STAT3-2 as a primary antibody against cell lysates. Relative expression of total STAT3 in SNC12 cells (A). Percentage of cells that express STAT3 (B). Average expression of STAT3 protein per cell (C). All data is normalized to β-tubulin expression. Click image to enlarge

CPTC-STAT3-2 Single Cell Western Blot (Cell Lysate)

Result: Positive

Single cell western blot using CPTC-STAT3-2 as a primary antibody against cell lysates. Relative expression of total STAT3 in SNC12 cells (A). Percentage of cells that express STAT3 (B). Average expression of STAT3 protein per cell (C). All data is normalized to β-tubulin expression.


Characterization SOP Files

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Antigen: Signal Transducer And Activator Of Transcription 3 Peptide 2
Background

NCI Identification Number:

00462

Antigen Name:

Signal Transducer And Activator Of Transcription 3 Peptide 2

CPTC Name:

CPTC-STAT3 Peptide 2

Aliases:

Signal Transducer And Activator Of Transcription 3; APRF; Acute-Phase Response Factor; Signal Transducer And Activator Of Transcription 3 (Acute-Phase Response Factor); DNA-Binding Protein APRF; ADMIO1; ADMIO; STAT3; HIES

Function:

The protein encoded by this gene 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. This protein is activated through phosphorylation in response to various cytokines and growth factors including IFNs, EGF, IL5, IL6, HGF, LIF and BMP2. This protein 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. The small GTPase Rac1 has been shown to bind and regulate the activity of this protein. PIAS3 protein is a specific inhibitor of this protein. Mutations in this gene are associated with infantile-onset multisystem autoimmune disease and hyper-immunoglobulin E syndrome. Alternative splicing results in multiple transcript variants encoding distinct isoforms. STAT3 (Signal Transducer And Activator Of Transcription 3) is a Protein Coding gene. Diseases associated with STAT3 include Hyper-Ige Recurrent Infection Syndrome 1, Autosomal Dominant and Autoimmune Disease, Multisystem, Infantile-Onset, 1. Among its related pathways are Common Cytokine Receptor Gamma-Chain Family Signaling Pathways and Interleukin-11 Signaling Pathway. Gene Ontology (GO) annotations related to this gene include DNA-binding transcription factor activity and sequence-specific DNA binding. An important paralog of this gene is STAT1.Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors (PubMed:10688651, PubMed:12359225, PubMed:12873986, PubMed:15194700, PubMed:17344214, PubMed:18242580, PubMed:23084476, PubMed:31462771). Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4 (PubMed:12873986). Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes (PubMed:12359225). Activated by IL31 through IL31RA (PubMed:15194700). 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 (PubMed:28065600). Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity (PubMed:23084476). Plays a crucial role in basal beta cell functions, such as regulation of insulin secretion (By similarity). STATs (signal transducers and activators of transcription) are a family of seven transcription factors that form part of the JAK-STAT signaling cascade, the basis of the signal transduction mechanism for many cytokine receptors. STATs are activated by phosphorylation by JAKs.

Chromosomal Localization:

17q21.2

Accession Number:

NP_644805.1

UniProt Accession Number:

P40763

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

YCRPESQEHPEADPGSAAPY(ph)LK

Native Sequence:

Calculated Isoelectric Point:

Molecular Weight:

2420

Last Updated:

11/19/2020

Links

Characterization Data

SOPs

No SOPs available.

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