Alkaline Phosphatase/ALPP Overexpression Lysate Summary
Description |
Alkaline Phosphatase/ALPP Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC210504
Accession#: NM_001632
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 |
ALPP |
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 |
55.5 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 Alkaline Phosphatase/ALPP Overexpression Lysate
Background
There are at least four distinct but related alkaline phosphatases: intestinal, placental, placental-like, and liver/bone/kidney (tissue non-specific). The first three are located together on chromosome 2 while the tissue non-specific form is located on chromosome 1. The product of this gene is a membrane bound glycosylated enzyme, also referred to as the heat stable form, that is expressed primarily in the placenta although it is closely related to the intestinal form of the enzyme as well as to the placental-like form. The coding sequence for this form of alkaline phosphatase is unique in that the 3' untranslated region contains multiple copies of an Alu family repeat. In addition, this gene is polymorphic and three common alleles (type 1, type 2 and type 3) for this form of alkaline phosphatase have been well characterized. [provided by RefSeq]. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.
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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Publications for Alkaline Phosphatase/ALPP Lysate (NBL1-07488) (0)
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Product General Protocols
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Video Protocols
FAQs for Alkaline Phosphatase/ALPP Lysate (NBL1-07488). (Showing 1 - 1 of 1 FAQ).
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I am looking to use a placental alkaline phosphatase antibody for sorting syncytiotrophoblast material from female tissue. It looks like most people who use these antibodies use in-house versions and references for commercially available antibodies I've found are from the '80's. I noticed you have several different monoclonal antibodies and was hoping you could give me a little more information on them and potential applications they've been tested for. Also, do you have a biotinylated or fluorophore-conjugated version?
- None of our placental alkaline phosphatase antibodies are biotinylated or fluorchrome conjugated but we offer a number of kits for this application and our lab can do a custom conjugation as well, for an additional fee. NB600-750 is useful in FACS with human cells, so I would recommend this antibody for your experiment. We back the use of this antibody as stated on the datasheet with the 100% Novus Guarantee. If you have questions about another antibody, I will be happy to answer them.
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