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Submit ReviewSPiDER-βGal Cellular Senescence Dye New
Description: Fluorogenic β-galactosidase substrate suitable for live/fixed cells and tissues imaging.
Chemical Name: 6'-(Diethylamino)-4'-(fluoromethyl)spiro[isobenzofuran-1(3H),9'-[9H]xanthen]-3'-yl-β-D-galactopyranoside
Purity: ≥85% (HPLC)
Biological Activity
Key information: SPiDER-βGal Cellular Senescence Dye is a fluorogenic β-galactosidase substrate suitable for labeling live cells in culture and living tissues. It exhibits activation of fluorescence upon reaction with the enzyme, remains inside cells by anchoring itself to intracellular proteins and provides single-cell resolution.
Used for: labeling and imaging lacZ reporters in living cells, tissues and model organisms. Also used to investigate senescence.
Application: Confocal microscopy, flow cytometry, fluorescence microscopy. SPiDER-βGal Cellular Senescence Dye has high sensitivity for the detection of peritoneal ovarian cancer metastases in animal models and displays high constrast of tumor margins. SPiDER-βGal Cellular Senescence Dye was used for investigating senescence of pancreatic cells and imaging adhesion G-protein-coupled receptors expressed in endothelial cells.
Properties and Photophysical Data: Excitation and emission maxima (λ) measured in 200 mM sodium phosphate buffer are 500-540 nm and 530-570 nm, respectively; activation ratio >650-fold.
Please refer to this protocol for further information on how to use this product.
Scientific Data
Fluorescence Imaging
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SPiDER-beta Gal labels beta -galactosidase-expressing cells in both live and fixed conditions. HEK/LacZ and HEK cells (1:1 co-culture) were incubated with 1 µmol/L SPiDER-beta Gal for 15 min at 37°C. (A) Living cells. (B) Cells fixed post-staining with 4% PFA/PBS. Fluorescence imaging: Ex 550/25 nm, Em 605/70 nm. Yellow: SPiDER-beta Gal; Blue: Hoechst 33342 (Catalog # 5117). Signal is retained following fixation, confirming compatibility with standard immunostaining workflows.
Flow Cytometry
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Flow cytometric detection of senescence-associated beta -galactosidase activity in WI-38 cells using SPiDER-beta Gal. WI-38 cells at passage 1 (grey) and passage 12 (green) were incubated with 1 µmol/L SPiDER-beta Gal for 15 min at 37°C and analysed by flow cytometry (Ex: 488 nm; Em: 500–600 nm). A marked rightward shift in fluorescence intensity at passage 12 reflects elevated SA-beta -gal activity consistent with replicative senescence. SPiDER-beta Gal enables quantitative discrimination of senescent cell populations by flow cytometry without the need for fixation.
Fluorescence Imaging
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Detection of senescence-associated beta -galactosidase (SA-beta -gal) activity using SPiDER-beta Gal. Cells were incubated with 1 µmol/L SPiDER-beta Gal for 30 min at 37°C and imaged by widefield fluorescence microscopy. Green: SPiDER-beta Gal (SA-beta -gal activity); Blue: Hoechst 33342 (Catalog # 5117). SPiDER-beta Gal self-immobilises to intracellular proteins following enzymatic cleavage, enabling high-contrast imaging of senescent cell populations.
Technical Data
| M. Wt | 567.6 |
| Formula | C31H34FNO8 |
| Storage | Store at +4°C |
| Purity | ≥85% (HPLC) |
| CAS Number | 1824699-57-1 |
| PubChem ID | 92135962 |
| InChI Key | SVEDIEUQTRCORD-JGANWENXSA-N |
| Smiles | O[C@H]([C@H](O)[C@H]1O)[C@@H](CO)O[C@H]1OC2=CC=C3C(OC(C=C(N(CC)CC)C=C4)=C4C35OCC6=C5C=CC=C6)=C2CF |
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
Product Datasheets
References
References are publications that support the biological activity of the product.
Doura et al (2016) Detection of lacZ-positive cells in living tissue with single-cell resolution. Angew.Chem.Int.Ed. 55 9620 PMID: 27400827
Nakamura et al (2017) A topically-sprayable, activatable fluorescent and retaining probe, SPiDER-βGal for detecting cancer: Advantages of anchoring to cellular proteins after activation. Oncotarget 8 39512 PMID: 28467810
Ibler et al (2019) Typhoid toxin exhausts the RPA response to DNA replication stress driving senescence and Salmonella infection. Nat.Commun. 10 4040 PMID: 31492859
If you know of a relevant reference for SPiDER-βGal Cellular Senescence Dye, please let us know.
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