DNMT3A Overexpression Lysate
Novus Biologicals, part of Bio-Techne | Catalog # NBL1-09970
Key Product Details
Species
Human
Applications
Western Blot
Product Summary for DNMT3A Overexpression Lysate
DNMT3A Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC213064
Accession#: NM_022552
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.
Expression Host: HEK293T
Plasmid: RC213064
Accession#: NM_022552
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.
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.
TMW
101.7 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.
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
Overexpression
Scientific Data Images for DNMT3A Overexpression Lysate
![Western Blot: DNMT3A Overexpression Lysate [NBL1-09970]](https://resources.bio-techne.com/images/products/Dnmt3a-Overexpression-Lysate-Adult-Normal-Western-Blot-NBL1-09970-img0002.jpg "Western Blot: DNMT3A Overexpression Lysate [NBL1-09970]")
Western Blot: DNMT3A Overexpression Lysate [NBL1-09970]
Western Blot: Dnmt3a Overexpression Lysate (Adult Normal) [NBL1-09970] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Dnmt3a.Formulation, Preparation, and Storage
Formulation
RIPA buffer
Concentration
The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.
Shipping
The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store at -80C. Avoid freeze-thaw cycles.
Background: DNMT3A
References
1. Thakur, A., Mackin, S. J., Irwin, R. E., O'Neill, K. M., Pollin, G., & Walsh, C. (2016). Widespread recovery of methylation at gametic imprints in hypomethylated mouse stem cells following rescue with DNMT3A2. Epigenetics Chromatin, 9, 53. doi:10.1186/s13072-016-0104-2
2. Ravichandran, M., Lei, R., Tang, Q., Zhao, Y., Lee, J., Ma, L., . . . Dawlaty, M. M. (2019). Rinf Regulates Pluripotency Network Genes and Tet Enzymes in Embryonic Stem Cells. Cell Rep, 28(8), 1993-2003.e1995. doi:10.1016/j.celrep.2019.07.080
3. Pang, Y., Liu, J., Li, X., Xiao, G., Wang, H., Yang, G., . . . Ren, H. (2018). MYC and DNMT3A-mediated DNA methylation represses microRNA-200b in triple negative breast cancer. J Cell Mol Med, 22(12), 6262-6274. doi:10.1111/jcmm.13916
4. Xia, L., Huang, W., Bellani, M., Seidman, M. M., Wu, K., Fan, D., . . . Baylin, S. B. (2017). CHD4 Has Oncogenic Functions in Initiating and Maintaining Epigenetic Suppression of Multiple Tumor Suppressor Genes. Cancer Cell, 31(5), 653-668.e657. doi:10.1016/j.ccell.2017.04.005
Long Name
DNA [Cytosine-5-]-Methyltransferase 3A
Alternate Names
DNA (cytosine-5-)-methyltransferase 3 alpha, DNA (cytosine-5)-methyltransferase 3A, DNA cytosine methyltransferase 3A2, DNA Methyltransferase 3 Alpha, DNA methyltransferase HsaIIIA, DNA MTase HsaIIIA, Dnmt3a, DNMT3A2, EC 2.1.1.37, M.HsaIIIA, TBRS
Gene Symbol
DNMT3A
Additional DNMT3A Products
Product Documents for DNMT3A Overexpression Lysate
Product Specific Notices for DNMT3A 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.
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