Apolipoprotein A-IV/ApoA4 Overexpression Lysate Summary
Description |
Apolipoprotein A-IV/ApoA4 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC222222
Accession#: NM_000482
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 |
APOA4 |
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 |
43.3 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 Apolipoprotein A-IV/ApoA4 Overexpression Lysate
Background
Apoliprotein (apo) A-IV gene contains 3 exons separated by two introns. A sequence polymorphism has been identified in the 3'UTR of the third exon. The primary translation product is a 396-residue preprotein which after proteolytic processing is secreted its primary site of synthesis, the intestine, in association with chylomicron particles. Although its precise function is not known, apo A-IV is a potent activator of lecithin-cholesterol acyltransferase in vitro.
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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Product General Protocols
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Video Protocols
FAQs for Apolipoprotein A-IV/ApoA4 Lysate (NBP2-09022). (Showing 1 - 1 of 1 FAQ).
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I would like to buy antibodies APOA4 (apolipoprotein AIV) for the cows (to western blot). I know that You don't have the antibodies for this species. Could You propose the best atibodies which I could test (use) in my experiment?
- All eight of our APOA4 antibodies were raised against the human protein, and all of them have currently only been tested against human samples. I ran an alignment for you and found that there is a 77% sequence homology between human and bovine APOA4, so it is possible that the antibodies raised against human APOA4 may recognise the bovine protein. Should you wish to try any of our antibodies with your samples, you may be interested in our Innovator's Reward Program, which allows you to try our primary antibodies in an untested species or application without the financial risk of failure. To participate you simply complete an online review with an image, detailing the positive or negative results of your study, and in return you receive a discount voucher for 100% of the purchase price of the reviewed product.
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