Human DMBT1 HEK293 Overexpression Lysate

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Human DMBT1 HEK293 Overexpression Lysate: Product Information

Product Description
This Human DMBT1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of DMBT1 protein (Cat: 11678-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the N-terminal segment of human DMBT1 (NP_004397.2) (Met 1-Ser 220) was expressed, fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human DMBT1 consists of 211 amino acids and has a predicted molecular mass of 22.6 kDa. The apparent molecular mass of rh DMBT1 is approximately 35-45 kDa in SDS-PAGE under reducing conditions due to glycosylation.

Human DMBT1 HEK293 Overexpression Lysate: Usage Guide

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 (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis 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.
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 200μL pure water and boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human DMBT1 HEK293 Overexpression Lysate: Alternative Names

Human GP340 Overexpression Lysate; Human muclin Overexpression Lysate

DMBT1 Background Information

Deleted in malignant brain tumors 1 protein, also known as glycoprotein 34, surfactant pulmonary-associated D-binding protein, DMBT1 and GP34, is a secreted protein which belongs to theDMBT1 family. DMBT1 contains 2CUB domains, 14SRCR domains and 1ZP domain. It is highly expressed in alveolar and macrophage tissues. In some macrophages, expression is detected on the membrane, and in other macrophages, it is strongly expressed in the phagosome/phagolysosome compartments. Defects in DMBT1 are involved in the development of glioma (GLM). Gliomas are central nervous system neoplasms derived from glial cells and comprise astrocytomas, glioblastoma multiforme, oligodendrogliomas , and ependymomas. DMBT1 may be considered as a candidate tumor suppressor for brain, lung, esophageal, gastric, and colorectal cancers. It may play roles in mucosal defense system, cellular immune defense and epithelial differentiation. DMBT1 may play a role as an opsonin receptor for SFTPD and SPAR in macrophage tissues throughout the body, including epithelial cells lining the gastrointestinal tract. It may be an important factor in fate decision and differentiation of transit-amplifying ductular (oval) cells within the hepatic lineage. DMBT1 may function as a binding protein in saliva for the regulation of taste sensation. It binds to HIV-1 envelope protein and has been shown to both inhibit and facilitate viral transmission.
Full Name
deleted in malignant brain tumors 1
References
  • Mollenhauer J., et al.,(1997), DMBT1, a new member of the SRCR superfamily on chromosome 10q25.3-q26.1 is deleted in malignant brain tumours. Nat. Genet. 17:32-39.
  • Holmskov U., et al., (1999), Cloning of gp-340, a putative opsonin receptor for lung surfactant protein D.Proc. Natl. Acad. Sci. U.S.A. 96:10794-10799.
  • Mollenhauer J., et al.,(1999), The genomic structure of the DMBT1 gene: evidence for a region with susceptibility to genomic instability.Oncogene 18:6233-6240.
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