ERp72 Knockout 293T Cell Lysate Summary
Preparation Method |
Knockout achieved by using CRISPR/Cas9,-22 bp deletion in exon3 and -5 bp deletion in exon3 and -2 bp deletion in exon3 |
Gene |
PDIA4 |
Applications/Dilutions
Dilutions |
|
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 293T Cell |
Life Stage |
Embryonic |
Tissue |
Kidney |
Notes
Powered by EDIGENE.
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 ERp72 Knockout 293T Cell Lysate
Background
ERp72 is an endoplasmic reticulum luminal protein that is both a stress protein and a member of the protein disulfide isomerase family of proteins. Sequence analysis of ERp72 shows that it shares sequence identity with PDI (protein disulphide isomerase) at three discrete regions, as well as containing the ER retention signal KEEL at its C-terminus. ERp72 is shown to have three copies of sequence believed to be CGHC-containing active sites, which are also found in PDI. These CGHC sites are critical sites that are involved in the protein disulphide isomerization process. These results suggest that ERp72 is a newly described member of the family of CGHC-containing proteins that functions as a molecular chaperone. This antibody was raised by a genetic immunization technique. Genetic immunization can be used to generate antibodies by directly delivering antigen-coding DNA into the animal, rather than injecting a protein or peptide (Tang et al. PubMed: 1545867; Chambers and Johnston PubMed 12910245; Barry and Johnston PubMed: 9234514). The animal's cells produce the protein, which stimulates the animal's immune system to produce antibodies against that particular protein. A vector coding for a partial fusion protein was used for genetic immunisation of a mouse and the resulting serum was tested in Western blot against an E.coli lysate containing that partial fusion protein. Genetic immunization offers enormous advantages over the traditional protein-based immunization method. DNA is faster, cheaper and easier to produce and can be produced by standard techniques readily amenable to automation. Furthermore, the antibodies generated by genetic immunization are usually of superior quality with regard to specificity, affinity and recognizing the native protein.
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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