CASE.EDU:    HOME | DIRECTORIES | SEARCH
case western reserve university

Center for Inherited Disorders of Energy Metabolism

 

PYRUVATE CARBOXYLASE

 

Method Description: Radioactive enzyme assay of pyruvate dependent fixation of 14CO2 following conversion of the oxalacetate product to citrate in the presence of excess citrate synthase and acetyl CoA. Linearity is established with respect to time and amount of sample (protein). Assay of citrate synthase (CS) is used as an internal control for mitochondrial content.

Specimen
Specimen Requirements
Shipping Requirements
Cultured Skin Fibroblasts

REQUIRES ADVANCE SCHEDULING

Two confluent T25 flasks filled to the neck with media in a non-vented flask and tape/parafilm shut. If several assays are ordered (more than 3), the shipment of an additional T25 flask is recommended.

Also the cell line MUST be MYCOPLASMA tested. If your institution is unable to test for mycoplasma, please tell us when you schedule a shipment or send them to Bionique to get tested.

Amt Required: 2T25 flasks

Schedule a shipment time and ship the 2T25 flasks at room temperature with enough packing material so flasks do not leak.

Please notify us if you are unable to ship the skin fibroblasts on your scheduled shipment time.

Skin Biopsy

Biopsy must be obtained under sterile conditions. Be careful to remove all betadine and alcohol and rinse with saline prior to obtaining the sample. Use a 3 or 4mm punch biopsy kit or take from the site of incision for a muscle biopsy.

Place biopsy in a container of sterile Hanks Balance Salt Solution (HBSS), sterile Ringer's Lactate, or other physiological solution.

Minimum Amt Required: a bx punch or a piece

Call us to let us know you'll be sending the skin biopsy and ship at room temperature with enough packing material so container does not leak.

 

Liver

After a biopsy/autopsy, the tissue specimen should be immediately quick-frozen in liquid nitrogen.

Minimum Amt Required: 50 mg

 

Ship on dry ice. If biopsy/autopsy is performed on a Friday, keep sample at -70°C or colder over the weekend and ship on Monday.

 

Samples Accepted: Monday - Friday, 8:30 AM - 5:00 PM

Turn-Around-Time:

2-3 weeks for liver

3-4 weeks for skin fibroblasts cultured from T25

7-10 weeks for culturing skin fibroblasts from a skin biopsy (skin biopsies grow at different rates and sometimes can take 3 months or longer)

CPT Codes for assay of fibroblasts or liver: 84999, 82657, 84157, 80502

CPT Codes for Cell Culture: 88233, 88240

CPT Code for Mycoplasma testing if your institution is unable to do this: 86738

Specimens can be sent via Federal Express, DHL, or UPS using Overnight Priority/Next Morning.

Special Instructions for skin fibroblasts: Ship T25 flasks with Mycoplasma results. Also save back-up flasks of skin fibroblast culture(s) at your institution until you have received the test results in the event of breakage of flasks or contamination during transport.

Future Testing on skin fibroblasts: We freeze down plugs on the cell line(s) you send to us, so if additional testing needs to be ordered in the future, we will have plugs here at CIDEM to initiate.

 

 

Return to TESTS