Annexin V Overexpression Lysate
Novus Biologicals | Catalog # NBL1-07565
![Western Blot: Annexin V Overexpression Lysate [NBL1-07565]](https://resources.rndsystems.com/images/products/Annexin-V-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-07565-img0002.jpg "Western Blot: Annexin V Overexpression Lysate [NBL1-07565]")
Key Product Details
Species
Applications
Product Summary for Annexin V Overexpression Lysate
Annexin V Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC205619
Accession#: NM_001154
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.
Product Specifications
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
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.
Type
Scientific Data Images for Annexin V Overexpression Lysate
Western Blot: Annexin V Overexpression Lysate [NBL1-07565]
Western Blot: Annexin V Overexpression Lysate (Adult Normal) [NBL1-07565] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Annexin V.Formulation, Preparation, and Storage
Formulation
Concentration
Shipping
Storage
Background: Annexin V
Alternate Names
Gene Symbol
Additional Annexin V Products
Product Documents for Annexin V Overexpression Lysate
Certificate of Analysis
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Product Specific Notices for Annexin V Overexpression Lysate
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.
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.
Related Research Areas
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
FAQs for Annexin V Overexpression Lysate
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Q: I am studying apoptosis in adherent cells. Can I trypsinize the cells when using Annexin V-FITC to stain the cells?
A: Adherent cells undergoing apoptosis automatically detach from the plate and may not require trypsinization. If there are attached cells, they may be trypsinized without affecting phosphatidylserine (PS).
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Q: I do not have access to flow cytometer in my building. Can I stain for Annexin V and propidium iodide (PI), fix the cells, then analyze them in a different facility?
A: Fixing the cells can affect cell permeability, enhancing the uptake of Annexin V and PI intracellularly. As a result, the profile of fixed cells may be different from unfixed cells, and the use of appropriate controls is recommended.
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Q: I am studying apoptosis in adherent cells. Can I trypsinize the cells when using Annexin V-FITC to stain the cells?
A: Adherent cells undergoing apoptosis automatically detach from the plate and may not require trypsinization. If there are attached cells, they may be trypsinized without affecting phosphatidylserine (PS).
-
Q: I do not have access to flow cytometer in my building. Can I stain for Annexin V and propidium iodide (PI), fix the cells, then analyze them in a different facility?
A: Fixing the cells can affect cell permeability, enhancing the uptake of Annexin V and PI intracellularly. As a result, the profile of fixed cells may be different from unfixed cells, and the use of appropriate controls is recommended.