{"id":312,"date":"2017-07-05T11:52:14","date_gmt":"2017-07-05T16:52:14","guid":{"rendered":"http:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/?page_id=312"},"modified":"2017-07-05T13:46:49","modified_gmt":"2017-07-05T18:46:49","slug":"experimental-resources","status":"publish","type":"page","link":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/resources\/experimental-resources\/","title":{"rendered":"Experimental Resources"},"content":{"rendered":"

Biophysical\/Structural Biology Technologies & Requirements<\/span><\/strong><\/p>\n

X-ray of apo Antibody<\/span><\/p>\n

Contact: Joel Harp<\/p>\n

e-mail: joel.harp@vanderbilt.edu<\/a><\/p>\n

phone: (615)936-3885<\/p>\n

Amount of Material: 1-5mg concentrated to >= 10 mg\/mL<\/p>\n

Purity of Material: >95%<\/p>\n

Comments: Provide in low ionic strength buffer, complete sequence, may require additional material for optimization, concentration depends on solubility and has ranged from 10 mg\/mL to 44.5 mg\/mL<\/p>\n

X-ray of Antibody Complex
\n<\/span><\/p>\n

Contact: Amritraj Patra<\/p>\n

e-mail: amritraj.patra@vanderbilt.edu<\/a><\/p>\n

phone: (615)600-1596<\/p>\n

Amount of Material: 1-5mg of antibody-antigen complex<\/p>\n

Purity of Material: Purified by size exclusion column and concentrated to ~10 mg\/mL<\/p>\n

Comments: Provide in low ionic strength buffer, complete sequence, may require additional material for optimization, concentration depends on solubility and has ranged from 10 mg\/mL to 44.5 mg\/mL<\/p>\n

HD\/X of apo Antibody
\n<\/span><\/p>\n

Contact: Marcus Nagel<\/p>\n

e-mail: mbmnagel@gmail.com<\/a><\/p>\n

phone: (615)569-8861<\/p>\n

Amount of Material: 5umol+ (or 250ug+)<\/p>\n

Purity of Material: 50% +<\/p>\n

Comments: Detailed information of mature sequence and PTMs, preferably little glycosylation, no aggregates, no detergents, ideal concentration is around 2mg\/mL<\/p>\n

HD\/X of Antibody Complex
\n<\/span><\/p>\n

Contact: Marcus Nagel<\/p>\n

e-mail: mbmnagel@gmail.com<\/a><\/p>\n

phone: (615)569-8861<\/p>\n

Amount of Material: each 5umol+ (or 500ug+)<\/p>\n

Purity of Material: 50% +<\/p>\n

Comments: Detailed information of mature sequence and PTMs, preferably little glycosylation, no aggregates, no detergents, ideal concentration is around 2mg\/mL<\/p>\n

Cross-linking of Antibody Complex
\n<\/span><\/p>\n

Contact: Marcus Nagel<\/p>\n

e-mail: mbmnagel@gmail.com<\/a><\/p>\n

phone: (615)569-8861<\/p>\n

Amount of Material: 15ug +<\/p>\n

Purity of Material: 30% +<\/p>\n

Comments: Detailed information of mature sequence and PTMs, preferably little glycosylation, AB and Antigen can be premixed, preferably amine-free buffer<\/p>\n

ITC of Antibody Complex
\n<\/span><\/p>\n

Contact: Alberto Cisneros<\/p>\n

e-mail: alberto.cisneros@vanderbilt.edu<\/a><\/p>\n

phone: (512)635-7261<\/p>\n

Amount of Material: Cell sample 5-20 uM, 300uL – between 100ug and 1mg depending on molecular weight of antigen; Syringe concentration 50-200uM – 180 to 700ug for Fab, 525ug to 2.1mg for IgG<\/p>\n

Purity of Material:<\/p>\n

Comments: Materials must undergo buffer exchange and must be as pure as possible<\/p>\n

EM of Antibody Complex
\n<\/span><\/p>\n

Contact: Amritraj Patra<\/p>\n

e-mail: amritraj.patra@vanderbilt.edu<\/a><\/p>\n

phone: (615)600-1596<\/p>\n

Amount of Material: 50-100 uL of ~1 mg\/mL of sample in commonly used biological buffer<\/p>\n

Purity of Material: Ideally sample should be as homogeneous as possible<\/p>\n

Comments: Some sample buffers may cause problems, including presence of detergent, phosphate, reducing agent or glycerol<\/p>\n

Octett of Antibody Complex
\n<\/span><\/p>\n

Contact: Alex Sevy<\/p>\n

e-mail: alexander.m.sevy@vanderbilt.edu<\/a><\/p>\n

phone: (615)343-8064<\/p>\n

Amount of Material: 20-100 uL each of antibody and antigen<\/p>\n

Purity of Material: >90%<\/p>\n

Comments: Ideally the antigen should be biotinylated.\u00a0 Amount of protein needed is highly dependent on affinity.\u00a0 Accurate KD can only be determined for antibodies with high off rate<\/p>\n

Wasatch SPR of Antibody Complex
\n<\/span><\/p>\n

Contact: Alex Sevy<\/p>\n

e-mail: alexander.m.sevy@vanderbilt.edu<\/a><\/p>\n

phone: (615)343-8064<\/p>\n

Amount of Material: 20-100 uL each of antibody and antigen<\/p>\n

Purity of Material: >90%<\/p>\n

Comments: Can be done label-free. Amount of protein needed is highly dependent on affinity.<\/p>\n

SAXS of Antibody Complex
\n<\/span><\/p>\n

Contact: Clayton Wandishin<\/p>\n

e-mail: clayton.m.wandishin@vanderbilt.edu<\/a><\/p>\n

phone: (703)568-1944<\/p>\n

Amount of Material: 24uL of sample at 1 mg\/mL, 24uL of sample at 3 mg\/mL, 24uL of sample at 5 mg\/mL, 48uL of sample buffer (2 x 24uL wells).\u00a0 This comes out to 216ug total for each sample<\/p>\n

Purity of Material: >95% with either SEC or dialysis preparation<\/p>\n

Comments: Buffer needs to be EXACT match, not an equivalent, and must contain more than 1M salt with <200mM preferred.\u00a0 2% glycerol is also recommended.<\/p>\n","protected":false},"excerpt":{"rendered":"

Biophysical\/Structural Biology Technologies & Requirements X-ray of apo Antibody Contact: Joel Harp e-mail: joel.harp@vanderbilt.edu phone: (615)936-3885 Amount of Material: 1-5mg concentrated to >= 10 mg\/mL Purity of Material: >95% Comments: Provide in low ionic strength buffer, complete sequence, may require additional material for optimization, concentration depends on solubility and has ranged from 10 mg\/mL to…<\/p>\n","protected":false},"author":5620,"featured_media":0,"parent":25,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"tags":[],"class_list":["post-312","page","type-page","status-publish","hentry"],"_links":{"self":[{"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/pages\/312","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/users\/5620"}],"replies":[{"embeddable":true,"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/comments?post=312"}],"version-history":[{"count":5,"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/pages\/312\/revisions"}],"predecessor-version":[{"id":318,"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/pages\/312\/revisions\/318"}],"up":[{"embeddable":true,"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/pages\/25"}],"wp:attachment":[{"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/media?parent=312"}],"wp:term":[{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/my.dev.vanderbilt.edu\/nextgenvaccines\/wp-json\/wp\/v2\/tags?post=312"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}