{"id":199,"date":"2022-05-16T17:29:01","date_gmt":"2022-05-16T17:29:01","guid":{"rendered":"http:\/\/chaletloree.com\/?page_id=199"},"modified":"2025-09-17T15:39:27","modified_gmt":"2025-09-17T15:39:27","slug":"protein","status":"publish","type":"page","link":"http:\/\/chaletloree.com\/index.php\/protein\/","title":{"rendered":"protein"},"content":{"rendered":"
\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t
\n\t\t\t\t
\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t
\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t
<\/p>\n

Buffers for protein extraction<\/strong><\/p>\n

– mild lysis buffer<\/a><\/p>\n

– RIPA<\/p>\n

– NETN<\/p>\n

Phosphatase and Protease inhibitors<\/a><\/p>\n

Pyrogallol Red Molybdate precipitation<\/a><\/p>\n

Immunoprecipitation (english) (french<\/a>)<\/p>\n

Affinity pulldown (tandem purification via SFB tag)<\/a><\/p>\n

WGA pulldown<\/a><\/p>\n

laminin overlay<\/p>\n

Preparation polyacrylamide gels\u00a0<\/strong><\/p>\n

– Bistris\u00a0buffer system<\/p>\n

– Tris-Glycin buffer system<\/p>\n

– Tris-Acetate buffer system<\/p>\n

Gel transfer<\/strong><\/p>\n

– wet transfer<\/a><\/p>\n

– semi-dry transfer<\/a><\/p>\n

Blue Native Gel electrophoresis<\/strong><\/p>\n

–\u00a0Variant A: Schaegger protocol<\/a><\/p>\n

– Variant B: Wittig protocol (english) (french<\/a>)<\/p>\n

Western Blot<\/strong><\/p>\n

general protocol (english<\/a>) (french<\/a>)<\/p>\n

Western Blot stripping<\/a><\/p>\n

Home-made enhanced chemoluminescence solution<\/a><\/p>\n

Akta liquid chromatography usage<\/a><\/strong><\/p>\n

Production of soluble protein from bacteria<\/p>\n

Purification of (HIS)6-tagged proteins<\/p>\n

Purification of GST-tagged proteins<\/p>\n

Self-made blue sepharose column<\/a><\/p>\n

Production of recombinant ApoAI<\/a><\/p>\n

Production of TEV protease<\/a><\/p>\n

Desalting using Sepharose minicolumns (Pernefsky)<\/a><\/p>\n

Protein quantification:<\/strong><\/p>\n

– BCA assay<\/p>\n

Coomassie staining (colloidal)<\/p>\n

Coomassie staining (classic)<\/a><\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

 <\/p>\n

<\/div>\n\t\t\t<\/div>\n\t\t\t<\/div>\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t\t\n\t\t\t<\/div>\n\t\t\t\t\n\t\t\t\t\n\t\t\t<\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":1,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_et_pb_use_builder":"on","_et_pb_old_content":"\n

Buffers for protein extraction<\/p>\n\n

- mild lysis buffer<\/a><\/p>\n\n

- RIPA<\/p>\n\n

- NETN<\/p>\n\n

Phosphatase and Protease inhibitors<\/a><\/p>\n\n

Pyrogallol Red Molybdate precipitation<\/a><\/p>\n\n

Immunoprecipitation (english) (french<\/a>)<\/p>\n\n

Affinity pulldown (tandem purification via SFB tag)<\/a><\/p>\n\n

WGA pulldown<\/a><\/p>\n\n

laminin overlay<\/p>\n\n

Preparation polyacrylamide gels <\/p>\n\n

- Bistris buffer system<\/p>\n\n

- Tris-Glycin buffer system<\/p>\n\n

- Tris-Acetate buffer system<\/p>\n\n

Gel transfer<\/p>\n\n

- wet transfer<\/a><\/p>\n\n

- semi-dry transfer<\/a><\/p>\n\n

Blue Native Gel electrophoresis<\/p>\n\n

- Variant A: Schaegger protocol<\/a><\/p>\n\n

- Variant B: Wittig protocol (english) (french<\/a>)<\/p>\n\n

Western Blot (english<\/a>) (french<\/a>)<\/p>\n\n

Western Blot stripping<\/a><\/p>\n\n

Home-made enhanced chemoluminescence solution<\/a><\/p>\n\n

Akta liquid chromatography usage<\/a><\/p>\n\n

Production of soluble protein from bacteria<\/p>\n\n

Purification of (HIS)6-tagged proteins<\/p>\n\n

Purification of GST-tagged proteins<\/p>\n\n

Self-made blue sepharose column<\/a><\/p>\n\n

Production of recombinant ApoAI<\/a><\/p>\n\n

Production of TEV protease<\/a><\/p>\n\n

Desalting using Sepharose minicolumns (Pernefsky)<\/a><\/p>\n\n

Protein determination:<\/p>\n\n

- BCA assay<\/p>\n\n

Coomassie staining (colloidal)<\/p>\n\n

Coomassie staining (classic)<\/a><\/p>\n\n

 <\/p>\n\n

 <\/p>\n\n

 <\/p>\n\n

 <\/p>\n\n

 <\/p>\n\n","_et_gb_content_width":"","footnotes":""},"class_list":["post-199","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/199","targetHints":{"allow":["GET"]}}],"collection":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/users\/1"}],"replies":[{"embeddable":true,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/comments?post=199"}],"version-history":[{"count":4,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/199\/revisions"}],"predecessor-version":[{"id":299,"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/pages\/199\/revisions\/299"}],"wp:attachment":[{"href":"http:\/\/chaletloree.com\/index.php\/wp-json\/wp\/v2\/media?parent=199"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}