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DNase I Cell Lysate | DNASE1 cell lysate

Human DNase I HEK293 Overexpression Lysate

Gene Names
DNASE1; DNL1; DRNI
Applications
Western Blot
Synonyms
DNase I; Human DNase I HEK293 Overexpression Lysate; Human DNASE1/Deoxyribonuclease I/DNL1 HEK293 Cell Lysate (WB positive control); deoxyribonuclease I; Human DNL1 Overexpression Lysate; Human DRNI Overexpression Lysate; DNASE1 cell lysate
Ordering
For Research Use Only!
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading buffer).
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Sequence Positions
271aa
Sequence
Met1-Lys282
Applicable Applications for DNASE1 cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.2. Re-dissolve the pellet using 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human DNASE1 (P24855) (Met1-Lys282) was expressed with a polyhistidine tag at the C-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for DNASE1 cell lysate
DNase1, also known as deoxyribonuclease I and DNL1, is a member of the DNase family. DNaseI is a nuclease that cleaves DNA preferentially at phosphodiester linkages adjacent to a pyrimidine nucleotide, yielding 5'-phosphate-terminated polynucleotides with a free hydroxyl group on position 3', on average producing tetranucleotides. DNaseI binds to the cytoskeletal protein actin. It binds actin monomers with very high (sub-nanomolar) affinity and actin polymers with lower affinity. Mutations in DNase1 gene have been associated with systemic lupus erythematosus (SLE), an autoimmune disease. DNase1 is used to treat the one of the symptoms of cystic fibrosis by hydrolyzing the extracellular DNA in sputum and reducing its viscosity.

This Human DNase I overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DNase I protein from the overexpression lysate was verified.
References
Shak S. et al., 1991, Proc Natl Acad Sci. 87 (23): 9188-92. Yasutomo K. et al., 2001, Nat Genet. 28 (4): 313-4. Hakkim A. et al., 2010, Proc Natl Acad Sci. 107 (21): 9813-8.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
UniProt Accession #
Molecular Weight
Calculated: 19kDa/31kDa
Observed: 37kDa
NCBI Official Full Name
Deoxyribonuclease-1
NCBI Official Synonym Full Names
deoxyribonuclease I
NCBI Official Symbol
DNASE1
NCBI Official Synonym Symbols
DNL1; DRNI
NCBI Protein Information
deoxyribonuclease-1; Dornase alfa; DNase I, lysosomal; human urine deoxyribonuclease I
UniProt Protein Name
Deoxyribonuclease-1
Protein Family
UniProt Gene Name
DNASE1
UniProt Synonym Gene Names
DNL1; DRNI; DNase I
UniProt Entry Name
DNAS1_HUMAN

NCBI Description

This gene encodes a member of the DNase family. This protein is stored in the zymogen granules of the nuclear envelope and functions by cleaving DNA in an endonucleolytic manner. At least six autosomal codominant alleles have been characterized, DNASE1*1 through DNASE1*6, and the sequence of DNASE1*2 represented in this record. Mutations in this gene have been associated with systemic lupus erythematosus (SLE), an autoimmune disease. A recombinant form of this protein is used to treat the one of the symptoms of cystic fibrosis by hydrolyzing the extracellular DNA in sputum and reducing its viscosity. Alternate transcriptional splice variants of this gene have been observed but have not been thoroughly characterized. [provided by RefSeq, Jul 2008]

Uniprot Description

DNASE1: Among other functions, seems to be involved in cell death by apoptosis. Binds specifically to G-actin and blocks actin polymerization. Defects in DNASE1 are a cause of susceptibility to systemic lupus erythematosus (SLE). A chronic, inflammatory and often febrile multisystemic disorder of connective tissue. It affects principally the skin, joints, kidneys and serosal membranes. It is thought to represent a failure of the regulatory mechanisms of the autoimmune system. Belongs to the DNase I family.

Protein type: Deoxyribonuclease; Motility/polarity/chemotaxis; Secreted; Secreted, signal peptide; EC 3.1.21.1

Chromosomal Location of Human Ortholog: 16p13.3

Cellular Component: extracellular region; nuclear envelope; nucleus

Molecular Function: protein binding; deoxyribonuclease I activity; actin binding; endodeoxyribonuclease activity

Biological Process: apoptosis; DNA catabolic process, endonucleolytic

Disease: Systemic Lupus Erythematosus

Research Articles on DNASE1

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Product Notes

The DNASE1 dnase1 (Catalog #AAA8115274) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 271aa. AAA Biotech's DNase I can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the DNASE1 dnase1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Lys28 2. It is sometimes possible for the material contained within the vial of "DNase I, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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