Streptolysin O (Hemolytic streptococcus), functional
01-53120 µg01-531-5
5 x 20 µg
Storage: Ship at 4℃ or -20℃, and store at -20℃ (For longer period, years, store at -80℃)
Product: Recombinant streptolysin O. Functional in membrane pore formation to introduce
molecules into living animal cells. The product was highly purified from E.coli
over-expressing SLO of Group C hemolytic streptococci (His6-tagged to the signal
peptide removed N-terminal of SLO). The specific activity is as high as 600,0001,000,000 hemolytic units (HU) /mg (depending on Lot).
Applications

1) Functional studies2).Reagent for membrane pore formation to introduce small-to-macromolecules into livingcells (For protocol refer to or other references; Walev I et al “Delivery of proteins intoliving cells by reversible membrane permeabilization with streptolysin-O.” PNAS 98:3185-3190 (2001) PMID: 11248053)
3).Antigen for the measurement of anti-streptolysin O antibody (ASO) (diagnostic reagent),
ELISA4) Western blotting, Dot blotting,5) Immuno-chromatography6) SDS-PAGE
Measurement of the activity: Definition of 1HU is activation of 50% hemolysis by incubating
3% sheep red blood cells at 37℃ for 30 min.
Purity: Over 98% by SDS-PAGE (see Fig.1)
Form: 1 mg/ml in PBS (-), 1 mM DTT, 50% glycerol, sterilized by filtration. No additive nor
carrier protein.
Inactivated SLO can be reactivated by thiol reagents such as 20 mM cysteine or 10mM DTT (Palmer M.Toxicon 39: 1681-1689 (2001) PMID: 11595631 )Background: Streptolysin O (SLO) is a membrane-damaging extracellular toxin produced byhemolytic streptococci. The membrane-damaging activity is measured by hemolysis ofred-blood cells. SLO is easily inactivated in the presence of oxygen but can be reactivatedby thiol compounds, so it is also called thiol-activated cytolysin . SLO is produced not only
by Group A hemolytic streptococci but also by Group C and Group G strains. The aminoacid sequences are highly conserved among them and their homology is over 98%.
Data Link : UniProtKB Q54114 ((TACY_STREQ)
Fig1. Purified SLO analysed by SDS-PAGE.
The SLO has molecular mass of 60.4 kDa.
This product has molecular mass of 64.5 kDa

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Fig.2 Introduction of fluorescein dextranofdifferent

molecularweights

intoresealed cells. A. HeLa cells were incubatedwith or without (2000 kDa dextran w/o SLO)
0.13 µg/ml SLO on ice for 5 min. After wash with
PBS three times, the cells were further withtransport buffer containing propidium iodide at
32°C for 5 min. Semi-intact HeLa cells wereincubated with 1.5 mg/ml L5178Y cytosol, anATP regenerating system, GTP, glucose, and 100µg/ml fluorescein-dextran of 3, 10, 40, 70, or
2000 kDa at 32°C for 15 min, and then wereresealed by treatment with 1 mM CaCl2 at 32°Cfor 5 min. After incubation with DMEM
supplemented with FCS for 30 min, the cells
were observed by confocal microscopy. Since thecells without SLO treatment did not contain thefluorescence of propidium iodide, differential
interference contrast (DIC) image was shown.
Bar = 10 µm. B. HeLa cells were treated as
described in A, were trypsinized, and weresubjected to flowcytometry. The histograms offluorescein

fluorescenceofdextran

withdifferent molecular weight in PI-positive cells
were shown. Data from Kano F. et al, PLoS
One. 2012;7(8):e44127.

References：This product has been used in the following publications.
1. Maeda, Y. et al. GPHR is a novel anion channel critical for acidification and functions ofthe Golgi apparatus. Nat. Cell Biol. 10: 1135-45 (2008) PMID: 18794847 Permeabilizationof cells.
2. Furukawa K. et al. Reduction-triggered fluorescent amplification probe for thedetection ofendogenous RNAs in living human cells. Bioconjug Chem. 2009 May
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20;20(5):1026-36. PMID: 19374406 Introduction of probes for RNA into permeabilized
human HL cells
3. Thiery J. et al. Perforin activates clathrin- and dynamin-dependent endocytosis, which isrequired for plasma membrane repair and delivery of granzyme B for granzyme-mediatedapoptosis. Blood 2010 115:1582-1593. PMID: 20038786 Promotion of endocytosis
4. Kano F. et al. Hydrogen peroxide depletes phosphatidylinositol-3-phosphate fromendosomes in a p38 MAPK-dependent manner and perturbs endocytosis. Biochim Biophys
Acta. 2011 May;1813(5):784-801. PMID: 21277337. Permeabilization of HeLa cells.
5. Potez S. et al. Tailored protection against plasmalemmal injury by annexins withdifferent Ca2+ sensitivities. J Biol Chem. 2011 May 20;286(20):17982-91. PMID:
21454475 Permeabilization of HEK cells.
6. Kano F. et al. A resealed-cell system for analyzing pathogenic intracellular events:
perturbation of endocytic pathways under diabetic conditions. PLoS
One. 2012;7(8):e44127. PMID: 22952896 Introduction of molecules into HeLa cells.
7. Imai A. et al. MADD/DENN/Rab3GEP functions as a guanine nucleotide exchange factorfor Rab27 during granule exocytosis of rat parotid acinar cells. Arch BiochemBiophys. 2013 Aug 1;536(1):31-7. PMID: 23702376. Introduction of antibody into cell
8. Gao N and Lehrman MA. Mannose-6-Phosphate: A Regulator of LLO Destruction. : InkaBrockhausen (ed.), Glycosyltransferases: Methods and Protocols, 2013, Springer, Methods
in Molecular Biology, vol. 1022, DOI 10.1007/978- Inka Brockhausen (ed.),Glycosyltransferases: 1-62703-465-4_20. Link:.springer.com/protocol/10.1007 Introductionof mannose-6-phosphate into living cells. (The authors specifically recommend
BioAcdemia streptolysin O for cell permeabilization)
9. Matsuto M et al. Reconstitution of the targeting of Rab6A to the Golgi apparatus insemi-intact HeLa cells: A role of BICD2 in stabilizing Rab6A on Golgi membranes and aconcerted role of Rab6A/BICD2 interactions in Golgi-to-ER retrograde transport. BiochimBiophys Acta. 2015 Oct;1853(10 Pt A):2592-609. PMID: 25962623 Introduction of protein(Rab6A) into permeabilized Hela cells.
10. Yasuga H. et al. Logic gate operation by DNA translocation through biological nanopores.
PLoS One. 2016 Feb 18;11(2):e0149667. PMID: 26890568 Nanopore formation in bilayer
lipid menmbrae.

11. Ojima K. et al. Myosin substitution rate is affected by the amount of cytosolic myosinin cultured muscle cells. Anim Sci J. 2017 Nov;88(11):1788-1793. PMID:28631391
Permeabilization of cells
12. Kano F. et al. Establishment and phenotyping of disease model cells created bycell-resealing technique. Sci Rep. 2017 Nov 9;7(1):15167.

PMID:2912317011.

Reversible permeabilization of plasma membrane
13. Watanabe H. et al. Analysis of Pore Formation and Protein Translocation Using
Large

BiologicalNanopores.

AnalChem. 2017

Nov

7;89(21):11269-11277.

PMID:28980803. Permeabilization for protein translocation
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https://www.bioacademia.co.jp/en/Material Safety Data Sheet

Revised on August 9, 2019

1. IDENTIFICATION OF THE SUBSTANCE/MIXTURE AND OF THE COMPANY/UNDERTAKING
1.1 Product identifiers
Product name: Streptolysin O, functional recombinant protein
Product code: 01-531
1.2 Relevant identified uses of the substance or mixture and uses advised against

For research use

only and not for use in diagnosis or in human.
1.3 Details of the supplier of the safety data sheet
Responsible Party
Company Name: BioAcademia Inc.
Address: North Buiding, Research Institute for Microbial Diseases, Osaka University, 3-1
Yamadaoka, Suita, Osaka 565-0871, Japan
Tel: 81-6-6877-2335

Fax: 81-6-6877-2336

E-mail: Info@bioacademia.co.jp
1.4 Emergency telephone number: 81-6-6877-2335

2. HAZARDS IDENTIFICATION
2.1 Classification of the substance or mixture
Not a hazardous substance or mixture according to Regulation (EC) No. 1272/2008. This substanceis not classified as dangerous according to Directive 67/548/EEC. 2.2 Label elements The productdoes not need to be labelled in accordance with EC directives or respective national laws.
2.2 Label elements The product does not need to be labelled in accordance with EC directives orrespective national laws.
2.3 Other hazards – none

3. COMPOSITION/INFORMATION ON INGREDIENTS
3.1 Composition of product: Streptolysin O (0.1%, CAS No. N/A), Glycerol (50%, CAS No. 56-81-5 ), x1
Phosphate Buffered Saline (KH2PO4 0.021%, CAS No. 7778-77-0 : NaCl 0.9%,CAS No. 7647-14-5:
Na2HPO4 0.038%, CAS No 7558-79-4.), Dithiothreitol (0.015%,CAS No. 3483-12-3), H2O (49%,CASNo. 7732-18-5)

4. FIRST AID MEASURESBioAcademia,Inc.

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4.1 Ingestion: Wash out with large amount of water. When swallowed, get medical attention if any
discomfort arises.
4.2 Eye contact: Wash with large amounts of water while lifting eye lids. Call medical doctor if
irritation continues.
4.3 Skin contact: Wash off with soap and plenty of water.
4.4 Spill release: Wear glove and sweep up the spill and then wash spill site. All the contaminants
should be autoclaved at 121℃ for 20 min before disposal.

5.Handling and Storage
5.1 Handling and Storage Precautions:BIOHAZARD. DO NOT USE IF SKIN IS CUTOR SCRATCHED.
5.2 Other Precautions:CAUTION: SUBSTANCE NOT YET FULLY TESTED.

6. Toxicological Information
6.1 Target: Cholesterol on human and animal cell membrane
6.2 Health Hazards: May be fatal if large amount enters bloodstream.
6.2 LD50 - Lethal dose (50 percent kill) intravenous,Rabbit, 1500 ng/kg (Ref :PHTHDT Pharmacology and Therapeutics.

(Pergamon PressLtd.,Headington Hill Hall, Oxford OX3 0BW, UK) Vol.(Issue) 11, Page 661,1981 )Guinea pig, 12 ug/kg (Ref: BICMBE Biochimie.

(SPPIF, B.P.22, F-41353 Vineuil, France,Vol.(Issue)55,Page 1187, 1973)
Toxicity is much less when introduced via other routes of entry like Interdermal injection

7. Exposure Controls/Personal Protection
7.1 Protective Gloves:COMPATIBLE CHEMICAL-RESISTANT GLOVES.
7.2 Eye Protection:ANSI APPROVED CHEMICAL WORKERS GOGGLES .
7.3 Other Protective Equipment:EYE WASH AND DELUGE SHOWER MEETING ANSIDESIGN CRITERIA .
7.3 Work Hygienic Practices:WASH THOROUGHLY AFTER HANDLING.

8. Disposal Considerations
Waste Disposal Methods: Autoclave the waste at 121℃ for 20 min.

9. Regulatory Information
Federal Regulatory Information:EUROPEAN INFORMATION: CAUTION: SUBSTANCE
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NOT YET FULLY TESTED.

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN
DIAGNOSTIC PROCEDURES. NOT FOR MILITARY USE.

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