Rabbit Anti-Amphiregulin antibody

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AR; Colorectum cell derived growth factor; CRDGF; A REG; AR antibody; AREG; Colorectum cell derived growth factor; CRDGF; MGC13647; Schwannoma derived growth factor; SDGF; AREG_HUMAN.

Cat:

SL3847R

Species Reactivity：

Human,Mouse,Rat,(predicted: Dog,Cow,Horse,Rabbit,Sheep,)

Immunogen：

KLH conjugated synthetic peptide derived from human Amphiregulin:185-252/252

Format：

Liquid

Storage instructions：

Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Concentration：

1mg/ml

Clonality：

Polyclonal

Isotype：

IgG

Applications：

WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500IF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.

Host：

Rabbit

Product Overview：

Sample: Lane 1: Mouse Placenta tissue lysates Lane 2: Mouse Breast tissue lysates Lane 3: Mouse Testis tissue lysates Lane 4: Rat Placenta tissue lysates Lane 5: Rat Breast tissue lysates Lane 6: Rat Testis tissue lysates Lane 7: Human MCF-7 cell lysates Lane 8: Human HepG2 cell lysates Lane 9: Human A431 cell lysates Primary: Anti- Amphiregulin (SL3847R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 16 kDaObserved band size: 47 kDaSample: HepG2(Human) Cell Lysate at 30 ugPrimary: Anti-Amphiregulin (SL3847R) at 1/2000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 26 kDObserved band size: 41 kDSample: A549(Human) Cell Lysate at 30 ugPrimary: Anti-Amphiregulin (SL3847R) at 1/500 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 26 kDObserved band size: 41 kDParaformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.Paraformaldehyde-fixed, paraffin embedded (human tonsil); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.Tissue/cell: mouse embryo tissue;4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Amphiregulin Polyclonal Antibody, Unconjugated(SL3847R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nucleiTissue/cell: mouse fetal liver;4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Amphiregulin Polyclonal Antibody, Unconjugated(SL3847R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nucleiTissue/cell: mouse fetal intestine;4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Amphiregulin Polyclonal Antibody, Unconjugated(SL3847R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nucleiBlank control:A549. Primary Antibody (green line): Rabbit Anti-Amphiregulin antibody (SL3847R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 20% PBST for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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Unit:

Price: $220.00

Product PDFs

Datasheet: Down Datasheet

Other Information
Citations
Protein Attributes
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Amphiregulin expression is induced by phorbol ester, estrogen, androgen, and other EGFR ligands. In vitro, amphiregulin functions as an autocrine growth stimulator to drive proliferation of colon carcinoma cells, normal and oncogene-transformed mammary epithelial cells, cervical carcinoma cells, prostate cancer cells, and keratinocytes. Amphiregulin is important in the development of human placenta and murine mammary gland. It is reportedly overexpressed in human cancers of breast, colon, stomach and pancreas.

Function:
Ligand of the EGF receptor/EGFR. Autocrine growth factor as well as a mitogen for a broad range of target cells including astrocytes, Schwann cells and fibroblasts.

Subunit:
The immature precursor interacts with CNIH.

Subcellular Location:
Membrane; Single-pass membrane protein.

Similarity:
Belongs to the amphiregulin family.
Contains 1 EGF-like domain.

SWISS:
P15514

Gene ID:
374

Database links:

Entrez Gene: 374 Human

Entrez Gene: 727738 Human

Entrez Gene: 11839 Mouse

Omim: 104128 Human

SwissProt: P15514 Human

SwissProt: P31955 Mouse

Unigene: 270833 Human

Unigene: 645475 Human

Unigene: 8039 Mouse

Picture

Sample:
HepG2(Human) Cell Lysate at 30 ug
Primary: Anti-Amphiregulin (SL3847R) at 1/2000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 41 kD

Sample:
A549(Human) Cell Lysate at 30 ug
Primary: Anti-Amphiregulin (SL3847R) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 26 kD
Observed band size: 41 kD

Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human tonsil); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human liver carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Amphiregulin) Polyclonal Antibody, Unconjugated (SL3847R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructions and DAB staining.

Tissue/cell: mouse embryo tissue;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Amphiregulin Polyclonal Antibody, Unconjugated(SL3847R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nuclei

Tissue/cell: mouse fetal liver;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Amphiregulin Polyclonal Antibody, Unconjugated(SL3847R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nuclei

Tissue/cell: mouse fetal intestine;4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Amphiregulin Polyclonal Antibody, Unconjugated(SL3847R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(SL0295G-Cy3)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,SLC0033) was used to stain the cell nuclei

Blank control:A549.
Primary Antibody (green line): Rabbit Anti-Amphiregulin antibody (SL3847R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 20% PBST for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Product Feedback Wall

Specific References (8) | SL3847R has been referenced in 8 publications.

[IF=2.424] Man She. et al. Dynamic Changes of AREG in the Sclera during the Development of Form-Deprivation Myopia in Guinea Pigs. 2021 Nov 12 WB,IF ; Guinea pig.

PubMed:34766531

[IF=3.998] Oka M et al. Dedifferentiation of smooth muscle cells in intracranial aneurysms and its potential contribution to the pathogenesis. Sci Rep. 2020 May 20;10(1):8330. IHC ; Human.

PubMed:32433495

[IF=0] Jiang, Wen Jun, et al. "Amphiregulin Antibody and Reduction of Axial Elongation in Experimental Myopia." EBioMedicine (2017). IHSLCF ; Guinea Pig.

PubMed:28251280

[IF=6.823] Bonnie Douglas. et al. Transgenic expression of a T cell epitope in Strongyloides ratti reveals that helminth-specific CD4+ T cells constitute both Th2 and Treg populations. Plos Pathog. 2021 Jul;17(7):e1009709 IF ; Rat.

PubMed:34237106

[IF=6.684] Huihua Kai. et al. LncRNA NORAD Promotes Vascular Endothelial Cell Injury and Atherosclerosis Through Suppressing VEGF Gene Transcription via Enhancing H3K9 Deacetylation by Recruiting HDAC6. Front Cell Dev Biol. 2021; 9: 701628 WB ; Human.

PubMed:34307380

[IF=1.314] Mukharram M. Bikbov. et al. Compatibility of intravitreally applied epidermal growth factor and amphiregulin. 2021 Mar 13 IHC ; Rabbit.

PubMed:33713254

[IF=3.998] Seung Seob Son. et al. In vivo silencing of amphiregulin by a novel effective Self-Assembled-Micelle inhibitory RNA ameliorates renal fibrosis via inhibition of EGFR signals. Sci Rep-Uk. 2021 Jan;11(1):1-14 IHC ; Mouse.

PubMed:33500443

[IF=4.6] c "Evaluation of estrogenic potency of a standardized hops extract on mammary gland biology and on MNU-induced mammary tumor growth in rats." The Journal of Steroid Biochemistry and Molecular Biology (2017). IHSLCP ; Rat.

PubMed:28964928