Lamin B2 Overexpression Lysate Summary
Description |
Lamin B2 Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC200807
Accession#: NM_032737
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 |
LMNB2 |
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 |
67.5 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 Lamin B2 Overexpression Lysate
Background
An important part of the cell nucleus is formed by nuclear lamina. Nuclear lamins form a network of filaments at the nucleoplasmic site of the nuclear membrane. Two main subtypes of nuclear lamins can be distinguished, i.e. A-type lamins and B-type lamins. The A-type lamins comprise a set of three proteins arising from the same gene by alternative splicing, i.e. Lamin A, Lamin C and lamin Adel10, while the B-type lamins include two proteins arising from two distinct genes, i.e. Lamin B1 and Lamin B2. The nuclear lamins comprise a unique subclass of the intermediate filament protein family. They share a molecular domain organisation with the other intermediate filament proteins in that they are fibrous molecules that have an aminoterminal globular head, a central rod of a-helices and a carboxyterminal globular domain. Many biochemical and molecular features of lamins have been studied, but their functions remain still largely undetermined. One of the functions ascribed to the lamina is the maintenance of the structural integrity of the nucleus. Besides interactions with the nuclear membrane and other intermediate filaments, lamins interact with the nuclear chromatin. Eukaryotic chromatin is organized into loops, which are attached to the nuclear matrix. This organization is thought to contribute to compaction of the chromatin and regulation of gene expression. Lamins, as part of the nuclear matrix, may be involved in these processes since chromatin binding sites have been detected in both A- and B-type lamins.
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 Lamin B2 Lysate (NBP2-05979). (Showing 1 - 1 of 1 FAQ).
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I'm looking for human lamin antibodies that might cross react with Drosophila lamins. I've found a few that I'm interested in, one of which comes in a smaller sample size. The other two do not have a smaller size ordering option. I was wondering if it would be possible to get these other antibodies in a smaller size as well, so we can test reactivity with Drosophila lamins before ordering the full-sized product?
- We have a wide range of antibodies to human Lamin. Unfortunately none of these has yet been tested against Drosophila samples, however you may be interested in our Innovator's Reward Program. This allows you to try our primary antibodies in an untested species or application, without the financial risk of failure. To participate you simply complete an online review with an image, detailing the positive or negative results of your study, and in return you receive a discount voucher for 100% of the purchase price of the reviewed product. We are unable to offer free samples of our antibodies but, as you rightly point out, some are available as smaller, sample size aliquots. If a sample size is not listed for a product I am afraid we are unable to offer a smaller aliquot than those already listed.
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