Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human, Mouse

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Cited:

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

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all Snail Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Snail
Pro2-Arg264
Accession # O95863

Specificity

Detects human Snail in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human Snail Antibody

Detection of Human Snail antibody by Western Blot.

Detection of Human Snail by Western Blot.

Western blot shows lysates of A549 human lung carcinoma cell line and JEG-3 human epithelial choriocarcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). A specific band was detected for Snail at approximately 29 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of Snail-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of Snail-regulated Genes by Chromatin Immunoprecipitation.

Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Snail/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. TheE-Cadherinpromoter was detected by standard PCR.

Snail antibody in A549 Human Cell Line by Immunocytochemistry (ICC).

Snail in A549 Human Cell Line.

Snail was detected in immersion fixed A549 human lung carcinoma cell line treated with Recombinant Human TGF-beta 1 (left panel, Catalog # 240-B) or untreated (right panel) using Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Snail antibody in A549 Human Cell Line antibody by Flow Cytometry.

Detection of Snail in A549 Human Cell Line by Flow Cytometry.

A549 human lung carcinoma cell line was stained with Goat Anti-Human Snail Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3639, filled histogram) or isotype control antibody (Catalog # AB-108-C, open histogram), followed by Fluorescein-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0109). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012). View our protocol for Staining Intracellular Molecules.

Detection of Human Snail by Western Blot

Detection of Human Snail by Western Blot

SerpinA1 was regulated by SnailA. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail and serpinA1 protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (serpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and western blot analysis was performed for detection of Snail and SerpinA1 expression. C. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail) or control vector pcDNA (vector), and ChIP assays were performed. The presence of the serpinA1 promoter (−516/−4) was verified in immunoprecipitates with either mouse IgG or anti-Snail antibodies, and assay inputs were analyzed using real-time PCR. The samples were loaded on agarose gels. D. Data show promoter enrichment in the anti-Snail immunoprecipitate relative to IgG. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26015410), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Snail by Western Blot

Detection of Human Snail by Western Blot

SerpinA1 was regulated by SnailA. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail and serpinA1 protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (serpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and western blot analysis was performed for detection of Snail and SerpinA1 expression. C. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail) or control vector pcDNA (vector), and ChIP assays were performed. The presence of the serpinA1 promoter (−516/−4) was verified in immunoprecipitates with either mouse IgG or anti-Snail antibodies, and assay inputs were analyzed using real-time PCR. The samples were loaded on agarose gels. D. Data show promoter enrichment in the anti-Snail immunoprecipitate relative to IgG. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26015410), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Snail by Western Blot

Detection of Human Snail by Western Blot

Snail and serpinA1 promoted tumor progression through fibronectinA. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (SerpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. C. DLD-1 and SW480 cells were transfected with pcDNA-fibronectin (Fibronectin), control vector pcDNA (vector), fibronectin siRNA (siFibronectin), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. D., E. Invasion and migration assays were performed using transfected cells. Representative data are shown for cells that invaded (top) and migrated (bottom) in the presence of 1% FBS. *P < 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26015410), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Snail by Western Blot

Detection of Human Snail by Western Blot

Snail and serpinA1 promoted tumor progression through fibronectinA. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (SerpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. C. DLD-1 and SW480 cells were transfected with pcDNA-fibronectin (Fibronectin), control vector pcDNA (vector), fibronectin siRNA (siFibronectin), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. D., E. Invasion and migration assays were performed using transfected cells. Representative data are shown for cells that invaded (top) and migrated (bottom) in the presence of 1% FBS. *P < 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26015410), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Snail by Western Blot

Detection of Human Snail by Western Blot

Snail and serpinA1 promoted tumor progression through fibronectinA. DLD-1 and SW480 cells were transfected with pcDNA-Snail (Snail), control vector pcDNA (vector), Snail siRNA (siSnail), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. B. DLD-1 and SW480 cells were transfected with pcDNA-serpinA1 (SerpinA1), control vector pcDNA (vector), serpinA1 siRNA (siSerpinA1), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. C. DLD-1 and SW480 cells were transfected with pcDNA-fibronectin (Fibronectin), control vector pcDNA (vector), fibronectin siRNA (siFibronectin), or nontargeting siRNA (siNT), and Snail, serpinA1, and fibronectin protein levels were evaluated by western blot analysis. D., E. Invasion and migration assays were performed using transfected cells. Representative data are shown for cells that invaded (top) and migrated (bottom) in the presence of 1% FBS. *P < 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26015410), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Snail by Western Blot

Detection of Human Snail by Western Blot

PDGF-AB induces Endo-MT through NF-kB-mediated Snail expression in ECs. a Human brain microvacular ECs were treated with glioma-CM. RNA was isolated and subjected to RNA-seq analysis. Left, heat map for expression of EMT-related transcriptional factors. Right, fold change of these transcriptional factors (n = 3, mean ± SEM). b, c ECs were treated with 100 ng/ml PDGF-AA, PDGF-AB, and PDGF-BB. b Cell lysates were immunoblotted. c RNA was isolated and analyzed by RT-PCR. Results were normalized with GAPDH levels (n = 3, mean ± SEM). d ECs were transfected with siRNAs targeting Snail or control scrambled sequence, and treated with PDGF-AB. Cell lysates were immunoblotted. e ECs were transfected with siRNAs targeting Erg-1, NF-kappa B, or control scrambled sequence, and treated with PDGF-AB. Cell lysates were immunoblotted. f ECs were treated with PDGF-AB for 2 h. Cells were analyzed by immunofluorescence. g, h ECs were treated with PDGF-AB or control medium for 8 h. Nuclear extracts were immunoprecipitated with anti-NF-kappa B antibody or IgG, and subjected to ChIP analysis with primers #1 and #2. g DNA was resolved by agarose electrophoresis, and imaged. Shown are representative results with primer #2. The arrow indicates the amplified DNA in Snail promoter. h Quantitative PCR analysis (n = 3, mean ± SEM). i ECs were transfected with siRNAs targeting Snail or control scrambled sequence, 4 days after treatment with Ki8751, followed by cell viability analysis (n = 3, mean ± SD) Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/30150753), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human Snail by Immunohistochemistry

Detection of Human Snail by Immunohistochemistry

Representative images of positive IHC staining in benign and malignant prostate tissue and a negative control Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/28877722), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Snail Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: PMA and calcium ionomycin treated Jurkat human acute T cell leukemia cell line chromatin, E-Cadherin promoter detected by standard PCR.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed A549 human lung carcinoma cell line treated with Recombinant Human TGF-beta 1 (Catalog # 240-B)

Intracellular Staining by Flow Cytometry

0.25 µg/106 cells
Sample: A549 human lung carcinoma cell line were fixed and permeabilized with FlowX FoxP3 Fixation & Permeabilization Buffer Kit (Catalog # FC012)

Western Blot

0.5 µg/mL
Sample: A549 human lung carcinoma cell line and JEG‑3 human epithelial choriocarcinoma cell line

Reviewed Applications

Read 5 reviews rated 4 using AF3639 in the following applications:

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Snail

Snail is predicted 29 kDa nuclear zinc finger transcriptional repressor that contains an N-terminal basic SNAG domain followed by three classical and one atypical zinc finger domains. During development, Snail is required for the establishment of left-right axis asymmetry. It also regulates the transcription of E-cadherin and other genes involved in epithelial-mesenchymal transitions during cancer progression. Human Snail shares 88% amino acid sequence identity with mouse and rat Snail.

Alternate Names

SLUGH2, SNAH, SNAI1

Entrez Gene IDs

6615 (Human); 20613 (Mouse); 116490 (Rat)

Gene Symbol

SNAI1

UniProt

O95863

Additional Snail Products

Product Documents for Human Snail Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Snail Antibody

For research use only

Citations for Human Snail Antibody

Customer Reviews for Human Snail Antibody (5)

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Customer Images

Showing  1 - 5 of 5 reviews Showing All
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  • Human Snail Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Human colorectal cancer cell
    Species: Human
    Verified Customer | Posted 10/24/2023
    SNAIL expression was checked with shCtr or sh-geneX to determine influences on EMT. A clear band was observed at around 30KD.
    Human Snail Antibody AF3639
  • Human Snail Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: A549 human lung carcinoma cell line
    Species: Human
    Verified Customer | Posted 10/17/2019
    Human Snail Antibody AF3639
  • Human Snail Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HCT-116 human colorectal carcinoma cell line, HeLa human cervical epithelial carcinoma cell line and PC-3 human prostate cancer cell line
    Species: Human
    Verified Customer | Posted 07/23/2018
    Total cell lysates from HCT-116, HeLa and PC-3 were subjected to western blot. PVDF membrane were probed with 1 um/ml Human Snail Antibody (AF3639). A specific band was detected for Snail at approximately 30 kDa. This experiment was conducted under reducing conditions.
    Human Snail Antibody AF3639
  • Human Snail Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Bladder cancer tissue
    Species: Human
    Verified Customer | Posted 12/10/2017
    Human Snail Antibody AF3639
  • Human Snail Antibody
    Name: Preethi Vijayaraj
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: mesenchymal cells
    Species: Human
    Verified Customer | Posted 10/18/2016
    Human Snail Antibody AF3639

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Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Human Snail Antibody

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  • Q: Can snail antibody AF3639 be used for electrophoresis motility shift assays (EMSA) instead of ChIP?

    A: For IP/Co-IP and ChIP the antibodies need to provide sufficient binding affinity for the antibodies to stay bound to the protein targets during the precipitation steps, whereas in EMSA the antibodies will only need to bind to the protein targets during the migration through the weak electrophoretic forces. Almost all, if not all, antibodies that bind to their protein targets with reasonable affinity and do not fall off from their protein targets when bound to the targeted, labeled DNA/RNA molecules during PAGE will be rendered in the upper shift in EMSA, provided the binding doesn't block the interaction between the protein targets and the nucleic acids.
     

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