{"id":758,"date":"2022-09-09T19:41:42","date_gmt":"2022-09-09T19:41:42","guid":{"rendered":"https:\/\/lab.research.sickkids.ca\/casca\/?page_id=758"},"modified":"2024-09-16T13:01:37","modified_gmt":"2024-09-16T13:01:37","slug":"custom-metal-conjugation-of-antibodies","status":"publish","type":"page","link":"https:\/\/lab.research.sickkids.ca\/casca\/custom-metal-conjugation-of-antibodies\/","title":{"rendered":"Custom Metal Conjugation of Antibodies"},"content":{"rendered":"

Antibody Conjugations for Your Panels<\/strong><\/h2>\n

Metal-tagged Abs specific for markers commonly used in mass cytometry are commercially available but are expensive since there is only a single vendor. To reduce costs and allow greater flexibility in experimental design, we perform custom metal conjugations of purified antibodies that you purchase or produce from hybridomas<\/strong>.<\/p>\n<\/div>

This service provides the most cost-effective way to build panels optimized for your specific experimental context. Add Abs (ideally validated to work in flow cytometry or on fixed tissue sections as per the protocol you will use) to our base immune profiling panels for mouse or human or start from scratch and design your own. You will need to provide us with 100-150 micrograms of purified, protein carrier free Ab for a single reaction).<\/p>\n

Contact Tina Chen<\/a> <\/strong>for the list of currently available metal tags and pricing.<\/p>\n

<\/h4>\n<\/div><\/div><\/div>

Conjugation Services<\/strong><\/h2>\n<\/div>

<\/i><\/i><\/span>Metal Choice and Panel Design Considerations<\/span><\/a><\/h4><\/div>
\n

For CyTOF panel design there are some things to consider as described here.<\/a><\/p>\n

Consult\u00a0 with a Field Application Specialist from Standard Biotools to help you design your panel (choose the metal tags for each Ab)<\/p>\n<\/div><\/div><\/div>

<\/i><\/i><\/span>Available Metal Tags<\/span><\/a><\/h4><\/div>
\n

Download available metal tags:<\/p>\n

CASCA-Helios_IMC_Conjugation_Channels<\/a><\/p>\n<\/div><\/div><\/div>

<\/i><\/i><\/span>Purified antibody guidelines<\/span><\/a><\/h4><\/div>
\n

Please\u00a0provide 100-150ug of purified IgG per single conjugation reaction of each antibody (Ab)<\/p>\n

Monoclonals are ideal, but affinity-purified polyclonal Abs can work<\/p>\n

Ab must be carrier-free (no BSA, hydrolyzed protein, gelatin etc.)<\/p>\n

Many vendors can supply carrier-free Ab but this may need to be requested specifically<\/p>\n

Ideal Ab concentration is 0.5-1 mg\/mL<\/p>\n

Some vendors supply Abs in “MaxPar Ready” format, which is carrier free and pre-filtered to remove low MW protein<\/p>\n

Ab should have been previously validated\u00a0for your desired application (eg. flow cytometry, IHC)<\/p>\n

    \n
  • For Hyperion analysis of FFPE sections, Ab needs to work with heat-induced Ag retrieval at pH 9<\/li>\n
  • For Hyperion analysis of frozen sections, Ab needs to work after fixation with 4% formaldehyde for 10-30′<\/li>\n<\/ul>\n<\/div><\/div><\/div><\/div><\/div><\/div><\/div><\/div><\/div>\n","protected":false},"excerpt":{"rendered":"","protected":false},"author":359,"featured_media":0,"parent":0,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-758","page","type-page","status-publish","hentry"],"yoast_head":"\nCustom Metal Conjugation of Antibodies | CASCA | SickKids<\/title>\n<meta name=\"description\" content=\"We perform custom metal conjugations of purified antibodies that you purchase or produce from hybridomas, a cost effective alternative for Metal-tagged Abs.\" \/>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/lab.research.sickkids.ca\/casca\/custom-metal-conjugation-of-antibodies\/\" \/>\n<meta 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