hnRNP A2B1 Overexpression Lysate Summary
Description |
hnRNP A2B1 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC219318
Accession#: NM_002137
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 |
HNRNPA2B1 |
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 |
35.8 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 hnRNP A2B1 Overexpression Lysate
Background
RNA polymerase II transcripts in the nucleus are in complex with several different proteins called heterogeneous nuclear ribonucleoproteins (hnRNPs). These proteins act in biological activities such as transcription, pre-mRNA processing, cytoplasmic mRNA ranslation, and turnover.2. hnRNPs can be isolated either by immunoprecipitation or by sucrose gradient fractionation of cell extracts. Isolated hnRNPs consist of protein groups named A to U and many of these protein groups consist of more than one isoform.3 The major steadystate proteins of the isolated hnRNP complex are the A1, A2, B1, B2, C1, and C2 with a range of molecular weights (34-43 kDa).3 hnRNP-A2/B1 proteins are located in the nucleus of most tissues cells; however, the hnRNP-A2 protein is found also in the cytoplasm of skin and esophagus cells. In rat brain, hnRNP-A2/B1 proteins are found in neurons in the cerebral cortices, hippocampal formation, olfactory regions, caudate-putamin, and the supraoptic ucleus of the hypothalamus.4 Expression of hnRNP-A2/B1 proteins was found also in lung development and its over expression correlated with lung cancer.5
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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