Q: For antibodies : STAT5b H00006777-M03; STAT 5a NBR2-00622; STAT3 NBP2-22471; STAT 6 H00006778-M01 - do these detect phosphorylated, unphosphorylated or both forms of STATs?

A: Antibodies H00006777-M03, NBR2-00622, NBP2-22471, and H00006778-M01 were raised to recognize total STAT proteins, they do not differentiate between any posttranslational modifications and will recognize both phosphorylated and non-phosphorylated protein.

STAT3 Antibody (9D8)

Novus Biologicals | Catalog # NBP2-22471

(1)

Citations (5)

Product Documents

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices
Research Areas

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey, Primate

Cited:

Human

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Knockdown Validated

Cited:

Western Blot, IF/IHC

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG_2A Clone # 9D8

View all STAT3 Primary Antibodies »

Product Specifications

Immunogen

Residues 655-770 from human recombinant protein expressed in bacteria

Reactivity Notes

Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2A

Scientific Data Images for STAT3 Antibody (9D8)

Western Blot: STAT3 Antibody (9D8) [NBP2-22471]

Western Blot: STAT3 Antibody (9D8) [NBP2-22471] - Human breast cancer cell MDA-MB-231 was treated with carboplatin for 72 hours and the expression of p-Stat3 at Y705 and total Stat3 were detected by western blot. From verified customer review.

Immunocytochemistry/ Immunofluorescence: STAT3 Antibody (9D8) [NBP2-22471]

Immunocytochemistry/Immunofluorescence: STAT3 Antibody (9D8) [NBP2-22471] - Analysis of STAT3 using anti-STAT3 (9D8) monoclonal antibody (shown in green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were then blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with a mouse monoclonal antibody recognizing STAT3, at a dilution of 1:100 for at least 1 hour at room temperature. Cells were then washed with PBS and incubated with DyLight 488 goat-anti-mouse secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye.

Immunohistochemistry-Paraffin: STAT3 Antibody (9D8) [NBP2-22471]

Immunohistochemistry-Paraffin: STAT3 Antibody (9D8) [NBP2-22471] - Biopsies of normal and cancer tissues.

Chromatin Immunoprecipitation: STAT3 Antibody (9D8) [NBP2-22471] - Analysis performed using cross-linked chromatin from rat hepatoma cells treated with insulin. IP performed using a multiplex microplate Matrix ChIP assay with STAT3 monoclonal antibody. Chromatin aliquots from cells were used per ChIP pull-down. Quantitative PCR data done in quadruplicate using 1ul of DNA in 2ul SYBR real-time PCR reactions containing primers to amplify exon-1 or exon-28 of LAMC1.Quantitation of immunoprecipitated chromatin is presented as signal relative to the total amount of input chromatin. Results represent the mean +/- SEM for three experiments. A schematic representation of the rat LAMC1 locus is shown; boxes represent exons (black boxes = translated regions, white boxes = untranslated regions), the zigzag line represents an intron, and the straight line represents upstream sequence. Regions amplified by LAMC1 primers are represented by black bars. Data courtesy of the Innovators Program.

Western Blot: STAT3 Antibody (9D8) [NBP2-22471]

Western Blot: STAT3 Antibody (9D8) [NBP2-22471] - Analysis of 25ug HepG2 total lysate.

Western Blot: STAT3 Antibody (9D8) [NBP2-22471]

Western Blot: STAT3 Antibody (9D8) [NBP2-22471] - Analysis of 25ug of various whole cell lysates.

Western Blot: STAT3 Antibody (9D8) [NBP2-22471]

Western Blot: STAT3 Antibody (9D8) [NBP2-22471] - Analysis of 25ug of U2-OS lysate from STAT3 SMART pool siRNA transfected or non-targeting control transfected U2-OS cells onto a 4-20% Tris-HCl polyacrylamide gel.

Immunoprecipitation: STAT3 Antibody (9D8) [NBP2-22471]

Immunoprecipitation: STAT3 Antibody (9D8) [NBP2-22471] - Analysis of STAT3 was performed on HepG2 cells. The antigen: antibody complex was formed by incubating 750ug whole cell lysate with 2ug of mouse monoclonal antibody recognizing STAT3 overnight on a rocking platform at 4C. The immune-complex was then captured on 50ul Protein A/G Plus Agarose. Captured immune-complexes were then washed extensively and proteins eluted with 5X Reducing Sample Loading Dye. Samples were then resolved on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to PVDF membrane and blocked with 5% Milk/TBS-0.1%Tween for at least 1 hour. Membranes were then probed with a mouse monoclonal antibody recognizing STAT3 at a dilution of 1:5000 overnight rotating at 4C. Membranes were washed in TBST and probed with Clean-blot IP detection reagent at a dilution of 1:2000 for at least one hour. Membranes were washed and chemiluminescent detection was performed using Super Signal West Dura.

Western Blot: STAT3 Antibody (9D8) [NBP2-22471] -

N-EV & H-EV treatment promote macrophage M2 polarization by delivering miR-21-5p that targets PTEN. a, western blot analysis of PTEN protein expression level in induced macrophages. H/i-miR-EV, monocytes were induced with the presence of EV secreted by miR-21-5p-inhibited, hypoxia pre-challenged MSCs; H-EV + i-miR, monocytes were transfected with miR-21-5p inhibitor-expressing vector before induction with the presence of H-EV. Macrophages induced without MSC-EV were used as negative control (NC). b, c, flow cytometry determining the percentage of CD163+CD206+ cells among total CD68+ cells after induction. N-EV + O/E PTEN or H-EV + O/E PTEN, monocytes were transfected with PTEN overexpressing vector before N-EV or H-EV treatment, respectively. d–f, western blot detecting Akt & STAT3 protein expression as well as their activating phosphorylation (p-Ser473 for Akt & p-tyr705 for STAT3) in macrophages after induction. g–i, ELISA evaluating IL-10, TGF-beta & VEGF-alpha in macrophage culture medium after induction. Macrophages induced with the presence of N-EV were used as negative control in b–i. Tukey’s test was used for statistical analysis. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001 Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30736829), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

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Applications for STAT3 Antibody (9D8)

Application

Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1-3 ul

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:1600

Immunohistochemistry-Paraffin

1:1600

Immunoprecipitation

2 ug

Western Blot

1:5000

Application Notes

WB from verified customer review.

Reviewed Applications

Read 1 review rated 4 using NBP2-22471 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS with 1 mg/ml BSA

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C. Avoid freeze-thaw cycles.

Background: STAT3

Stat3 is a member of a group of proteins (Signal Transduction and Activators of Transcription) that act as both signal transducers for cytokines and growth factors, as well as transcription factors (1,2). Stats are activated by tyrosine phosphorylation in response to different ligands, after which they translocate to the cell nucleus (1). Stat3 is constitutively activated in a number of human tumors (3). Stat3 is activated by phosphorylation on tyrosine 705 which induces dimerization, nuclear translocation, and DNA binding While, Stat3-activated transcription seems to be regulated by serine phosphorylation at serine 727 (4)

Long Name

Signal Transducer and Activator of Transcription 3

Alternate Names

Acute-phase response factor, APRFMGC16063, DNA-binding protein APRF, FLJ20882, HIES, signal transducer and activator of transcription 3, signal transducer and activator of transcription 3 (acute-phase responsefactor)

Entrez Gene IDs

6774 (Human)

Gene Symbol

STAT3

UniProt

P40763

Additional STAT3 Products

Product Documents for STAT3 Antibody (9D8)

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Certificate of Analysis

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Product Specific Notices for STAT3 Antibody (9D8)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Apoptosis Transcription Factors and Regulators

Dendritic Cell Development

GDNF Family

Interferons

Myeloid-derived Suppressor Cells (MDSC)

PDGF Family

Transcription Factors in the Akt Pathway

Citations for STAT3 Antibody (9D8)

Customer Reviews for STAT3 Antibody (9D8) (1)

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STAT3 Antibody (9D8)

Name: Anonymous

Application: Western Blot

Sample Tested: Breast cancer cells

Species: Human

Verified Customer | Posted 10/11/2018

Human breast cancer cell MDA-MB-231 was treated with carboplatin for 72 hours and the expression of p-Stat3 at Y705 and total Stat3 were detected by western blot.

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Protocols

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FAQs for STAT3 Antibody (9D8)

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Q: For antibodies : STAT5b H00006777-M03; STAT 5a NBR2-00622; STAT3 NBP2-22471; STAT 6 H00006778-M01 - do these detect phosphorylated, unphosphorylated or both forms of STATs?

A: Antibodies H00006777-M03, NBR2-00622, NBP2-22471, and H00006778-M01 were raised to recognize total STAT proteins, they do not differentiate between any posttranslational modifications and will recognize both phosphorylated and non-phosphorylated protein.