Phospholipase A2 XII Overexpression Lysate Summary
Description |
Phospholipase A2 XII Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC202955
Accession#: NM_030821
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 |
PLA2G12A |
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 |
20.9 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 Phospholipase A2 XII Overexpression Lysate
Background
Secreted phospholipase A2 (sPLA2) enzymes liberate arachidonic acid from phospholipids for production of eicosanoids and exert a variety of physiologic and pathologic effects. Group XII sPLA2s, such as PLA2G12A, have relatively low specific activity and are structurally and functionally distinct from other sPLA2s (Gelb et al., 2000 [PubMed 11031251]).[supplied by OMIM]
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 Phospholipase A2 XII Lysate (NBP2-06175). (Showing 1 - 2 of 2 FAQ).
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Our customer asked about the cross reactivity of PLA2G12A antibodies. Do you have any info if these antibodies?
- Unless specifically indicated on the datasheet they are not tested to recognize other phospholipases. The immunogen information is present for all of those so that might give some clues to the customer as to whether they might have some reactivity, but as far as direct testing is concerned on the other phospholipases this has not been performed by the lab.
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Our customer asked about the cross reactivity of PLA2G12A antibodies. Do you have any info if these antibodies recognize other phospholipases?
- Unless specifically indicated on the datasheet they are not tested to recognize other phospholipases. The immunogen information is present for all of those so that might give some clues to the customer as to whether they might have some reactivity, but as far as direct testing is concerned on the other phospholipases this has not been performed by the lab.
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