Line 4: |
Line 4: |
| β-D-galactopyranoside, C19H21O9Na, MW 416.35, MDL Number MFCD03458481. | | β-D-galactopyranoside, C19H21O9Na, MW 416.35, MDL Number MFCD03458481. |
| | | |
− | S-Gal®, sodium salt is a patented water-soluble, autoclavable chromogenic substrate for β-galactosidase that is designed to replace X-Gal in blue-white selection of recombinant bacterial colonies with the lac+ phenotype. Caution: For black color development to occur, ferric ion must be present. Although we recommend ferric ammonium citrate (500 mg/L of media), other ferric compounds can be used to provide this requirement, depending upon your particular system. From [http://www.sigmaaldrich.com/catalog/ProductDetail.do?N4=S7313%7CSIGMA&N5=SEARCH_CONCAT_PNO%7CBRAND_KEY&F=SPEC Sigma-Aldrich site]. | + | S-Gal®, sodium salt is a patented water-soluble, autoclavable chromogenic substrate for β-galactosidase that is designed to replace X-Gal in blue-white selection of recombinant bacterial colonies with the lac+ phenotype. Caution: For black color development to occur, ferric ion must be present. Although we recommend ferric ammonium citrate (500 mg/L of media), other ferric compounds can be used to provide this requirement, depending upon your particular system. From [https://www.sigmaaldrich.com/US/en/product/sigma/s7313 Sigma-Aldrich site]. |
| | | |
| | | |
Line 22: |
Line 22: |
| ==S-Gal staining conditions== | | ==S-Gal staining conditions== |
| | | |
− | According to [http://www.sigmaaldrich.com/catalog/ProductDetail.do?N4=S7313%7CSIGMA&N5=SEARCH_CONCAT_PNO%7CBRAND_KEY&F=SPEC Sigma-Aldrich site]: | + | According to [https://www.sigmaaldrich.com/US/en/product/sigma/s7313 Sigma-Aldrich site]: |
| | | |
| Stock solutions of S-Gal can be made by dissolving at 50 mg/mL in deionized water, sterile-filtering and storing at -20 °C. | | Stock solutions of S-Gal can be made by dissolving at 50 mg/mL in deionized water, sterile-filtering and storing at -20 °C. |
Line 54: |
Line 54: |
| | | |
| | | |
− | http://www.bio.davidson.edu/courses/genomics/method/xgal.gif
| + | https://bio.davidson.edu/courses/genomics/method/xgal.gif |
| | | |
| Cleavage by beta-galactosidase yields a dihalogen-substituted indoxyl, 5-bromo-4-chloro-3-indoxyl: | | Cleavage by beta-galactosidase yields a dihalogen-substituted indoxyl, 5-bromo-4-chloro-3-indoxyl: |
| | | |
| Oxidation in the presence of an electron acceptor yields a relatively insoluble indigoid dye: | | Oxidation in the presence of an electron acceptor yields a relatively insoluble indigoid dye: |
− |
| |
− |
| |
| | | |
| =Oxidants used in histochemistry= | | =Oxidants used in histochemistry= |
Line 70: |
Line 68: |
| Performic acid PMID: 1099403 | | Performic acid PMID: 1099403 |
| | | |
− | Cepko, C., Ryder, E., Fekete, D.M., and Bruhn, S. Detection of B-galactosidase and alkaline phosphatease activities in tissue. Online: http://genepath.med.harvard.edu/~cepko/protocol/xgalplap-stain.htm. An excellent brief discussion of X-Gal histochemistry. Much of the material presented here was derived using their references. | + | Cepko, C., Ryder, E., Fekete, D.M., and Bruhn, S. Detection of B-galactosidase and alkaline phosphatease activities in tissue. Online (archive): https://web.archive.org/web/20160821210733/http://genepath.med.harvard.edu/~cepko/protocol/xgalplap-stain.htm. An excellent brief discussion of X-Gal histochemistry. Much of the material presented here was derived using their references. |
| | | |
| =Studies in Enzyme Cytochemistry= | | =Studies in Enzyme Cytochemistry= |
Line 87: |
Line 85: |
| | | |
| Recommends ferrocyanide, and notes that it inhibits red blood cell staining "presumably because of the chemical similarity to cyanide" | | Recommends ferrocyanide, and notes that it inhibits red blood cell staining "presumably because of the chemical similarity to cyanide" |
| + | |
| + | |
| + | [[Category:Taste Aversion]] |
| + | [[Category:c-Fos]] |
| + | [[Category:Protocols]] |