Moesin Knockout HeLa Cell Lysate

Product Discontinued

Moesin Knockout HeLa Cell Lysate Summary

• Preparation Method: Knockout achieved by using CRISPR/Cas9,1 bp insertion in exon 5
• Gene: MSN

Applications/Dilutions

• Dilutions:
  • Western Blot
• Application Notes: You will receive 1 vial (100ug) of knockout cell lysate and 1 vial (100ug) of Parental cell lysate. Lysate can be diluted with 1X SDS sample buffer and will be stable at -20 degrees C for 12 months. Minimize freeze-thaw cycles.

Packaging, Storage & Formulations

• Storage: Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.
• Buffer: 0.1 mg cell homogenate lyophilized in RIPA buffer made with double-knockout cell lines.
• Concentration: LYOPH
• Reconstitution Instructions: To use as WB negative control, spin down briefly and resuspend in 100 uL 1xSDS sample buffer (2% SDS, 60 mM Tris-HCl pH 6.8, 10% Glycerol, 0.02% Bromophenol blue, 60 mM beta-mercaptoethanol). Boil the lysate for 3 - 5 minutes before loading it onto gel.

Lysate Details for Array

• Type: Knockout HeLa Cell

Notes

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Validation of antibody specificity is critical and verification of antibody performance against knockout samples is one way to guarantee that an antibody recognizes a specific target. Novus' KO cell lysate can be used as a negative control for western blots and to confirm the specificity of antibodies.

Alternate Names for Moesin Knockout HeLa Cell Lysate

• Membrane-organizing extension spike protein
• moesin

Background

Moesin (for membrane-organizing extension spike protein) is a member of the ERM family which includes ezrin and radixin. ERM proteins appear to function as cross-linkers between plasma membranes and actin-based cytoskeletons. Moesin is localized to filopodia and other membranous protrusions that are important for cell-cell recognition and signaling and for cell movement. [provided by RefSeq, Jul 2008]

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.

FAQs for Moesin Lysate (NBP2-64964). (Showing 1 - 1 of 1 FAQ).

1. I am looking to use shRNA to inhibit Moesin expression. I have had people advise me that my initial MOI should be low as 'less is more' and 'a little goes a long way' in terms of siRNA. I was wondering if you could elaborate on this for me and explain why my initial MOI should be low.
   • The reason for a low MOI is most likely because RNAi is a very strong and efficient technique. Wikipedia does a good job of explaining <a href="http://en.wikipedia.org/wiki/RNA_interference" target="_blank">RNA interference</a>. However, I would imagine that in a cell, there will be at most 1-2 copies of the gene mRNA present at any given time, unless you're dealing with a highly expressed protein such as Actin, where I would imagine silencing Actin would be lethal to the cell. I can imagine a few reasons to not use too much siRNA. First, it is expensive, so you don't want to waste it. Second, using too much would cause there to be a lot of non-translatable RNA present in the cell, which could trigger an immune response, as the presence of uncapped RNAs can indicate presence of a virus and one of the TLRs may respond to this.

Bioinformatics

• Gene Symbol: MSN