Carbonic Anhydrase I/CA1 Overexpression Lysate Summary
Description |
Carbonic Anhydrase I/CA1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC225348
Accession#: NM_001128829
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
Gene |
CA1 |
Applications/Dilutions
Dilutions |
|
Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
Theoretical MW |
28.7 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
Packaging, Storage & Formulations
Storage |
Store at -80C. Avoid freeze-thaw cycles. |
Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for Carbonic Anhydrase I/CA1 Overexpression Lysate
Background
Carbonic anhydrase performs an important role in respiration by facilitating transfer of CO2. Carbonic Anhydrase I (CAI) is a single polypeptide chain metalloenzyme of molecular weight 29 kDa. It is present mainly in erythrocytes at a concentration of about 12 mg per gram haemoglobin. Its main enzyme function is to catalyse the reversible hydration of carbon dioxide. It also catalyses the hydrolysis of certain esters. The twelve active CA isozymes are thought to regulate a variety of cellular functions including several processes in the reproductive systems. A mutational deficiency has been described but patients exhibit no adverse chemical symptoms. Red cell CAI levels are decreased in patients with thyrotoxicosis.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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