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Purified Mouse Anti-SNAI2/Slug
Purified Mouse Anti-SNAI2/Slug

Immunoflourescent, flow cytometric, western blot, and immunohistochemical analysis of SNAI2/Slug in human sarcoma cells.

FAR LEFT: RH-30 rhabdomyosarcoma cells (DSMZ, ACC 489) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Perm/Wash™ Buffer (Cat No. 554723) and stained with Purified Mouse Anti-SNAI2/Slug (pseudo-colored green) at 1.2 μg/mL followed by Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies). Nuclear counter-staining was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software. Permeabilization with BD Phosflow™ Perm Buffer III (Cat. No. 558050) or 0.1% Triton™ X-100 buffer is also suitable.

MIDDLE LEFT: RH-30 cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574) and stained with either Purified Mouse IgG1 κ Isotype Control (Cat No. 554121, dashed line) or Purified Mouse Anti-SNAI2/Slug (solid line), followed by PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer.

MIDDLE RIGHT: Lysates of RH-30 cells were electrophoresed (SDS-PAGE), transferred to PVDF membranes and then probed with 125, 60, 30, 15 and 7 ng/mL of Purified Mouse Anti-SNAI2/Slug (lanes 1-5) and HRP goat anti-mouse Ig (Cat. No. 554002).

FAR RIGHT: Following antigen retrieval with Retrievagen A Buffer (Cat. No. 550524), formalin-fixed paraffin sections of a human fibrosarcoma were stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No.550878, left) or Purified Mouse Anti-SNAI2/Slug (right). Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) were used to develop the staining. Original magnification: 40×.

Immunoflourescent, flow cytometric, western blot, and immunohistochemical analysis of SNAI2/Slug in human sarcoma cells.

FAR LEFT: RH-30 rhabdomyosarcoma cells (DSMZ, ACC 489) were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), permeabilized with BD Perm/Wash™ Buffer (Cat No. 554723) and stained with Purified Mouse Anti-SNAI2/Slug (pseudo-colored green) at 1.2 μg/mL followed by Alexa Fluor® 488 goat anti-mouse Ig (Life Technologies). Nuclear counter-staining was with DAPI (pseudo-colored blue). The images were captured on a BD Pathway™ 435 Cell Analyzer and merged using BD Attovision™ software. Permeabilization with BD Phosflow™ Perm Buffer III (Cat. No. 558050) or 0.1% Triton™ X-100 buffer is also suitable.

MIDDLE LEFT: RH-30 cells were fixed and permeabilized with the BD Pharmingen™ Transcription Factor Buffer Set (Cat No. 562574) and stained with either Purified Mouse IgG1 κ Isotype Control (Cat No. 554121, dashed line) or Purified Mouse Anti-SNAI2/Slug (solid line), followed by PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat No. 550589). The fluorescence histograms were derived from gated events with the forward and side light-scatter characteristics of viable cells. Flow cytometry was performed using a BD LSRFortessa™ Cell Analyzer.

MIDDLE RIGHT: Lysates of RH-30 cells were electrophoresed (SDS-PAGE), transferred to PVDF membranes and then probed with 125, 60, 30, 15 and 7 ng/mL of Purified Mouse Anti-SNAI2/Slug (lanes 1-5) and HRP goat anti-mouse Ig (Cat. No. 554002).

FAR RIGHT: Following antigen retrieval with Retrievagen A Buffer (Cat. No. 550524), formalin-fixed paraffin sections of a human fibrosarcoma were stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No.550878, left) or Purified Mouse Anti-SNAI2/Slug (right). Biotin Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550337), Streptavidin HRP (Cat. No.550946), and the DAB Substrate Kit (Cat. No. 550880) were used to develop the staining. Original magnification: 40×.

Product Details
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BD Pharmingen™
Snai2; SLUG; Slugh; SLUGH
Human (QC Testing), Mouse (Tested in Development)
Mouse IgG1, κ
Human SNAI2 Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested), Bioimaging, Immunofluorescence, Immunohistochemistry-formalin (antigen retrieval required), Western blot (Tested During Development)
30-34 kDa
0.5 mg/ml
AB_2738865
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  4. Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
  5. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  6. Triton is a trademark of the Dow Chemical Company.
  7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
564614 Rev. 1
Antibody Details
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S43-1259

The S43-1259 monoclonal antibody specifically recognizes SNAI2, also named Slug, which is a zinc finger transcriptional repressor first recognized for its roles in embryogenesis. SNAI2/Slug and other members of the snail family of transcription factors inhibit the expression of E-cadherin, which plays important roles in cell adhesion and in the maintenance of tissue structure. SNAI2/Slug activation and E-cadherin inhibition induces epithelial-mesenchymal transition (EMT) at several stages of embryonic development and also contributes to the invasiveness of malignancies. Aberrant expression of SNAI2/Slug has also been linked to cancer stem cell formation, cell cycle regulation and apoptosis.  SNAI2/Slug is a downstream effector of various signaling pathways, including Akt, Wnt and SCF/ckit signaling.

564614 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
564614 Rev.1
Citations & References
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Development References (4)

  1. Carpenter RL, Paw I, Dewhirst MW, Lo HW. Akt phosphorylates and activates HSF-1 independent of heat shock, leading to Slug overexpression and epithelial-mesenchymal transition (EMT) of HER2-overexpressing breast cancer cells. Oncogene. 2014; ePub(Biology). View Reference
  2. Francí C, Takkunen M, Dave N, et al. Expression of Snail protein in tumor-stroma interface. Oncogene. 2006; 25(37):5134-5144. (Biology). View Reference
  3. Guo W, Keckesova Z, Donaher JL, et al. Slug and Sox9 cooperatively determine the mammary stem cell state. Cell. 2012; 148(5):1015-1028. (Biology). View Reference
  4. Hay ED, Zuk A. Transformations between epithelium and mesenchyme: normal, pathological, and experimentally induced. Am J Kidney Dis. 1995; 26(4):678-690. (Biology). View Reference
View All (4) View Less
564614 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.