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DGK-beta Overexpression Lysate

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Western Blot: DGKB Overexpression Lysate (Adult Normal) [NBP2-07914] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DGKB.
Western Blot: DGKB Overexpression Lysate (Adult Normal) [NBP2-07914] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DGKB.

Product Details

Summary
Product Discontinued
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Order Details


    • Catalog Number
      NBP2-07914
    • Availability
      Product Discontinued

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DGK-beta Overexpression Lysate Summary

Description
Transient overexpression lysate of diacylglycerol kinase, beta 90kDa (DGKB), transcript variant 1 The lysate was created in HEK293T cells, using Plasmid ID RC217461 and based on accession number NM_004080. The protein contains a C-MYC/DDK Tag. 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
DGKB

Applications/Dilutions

Dilutions
  • Western Blot
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
90.4 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

Type
Overexpression

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.This product is manufactured by and sold under license from OriGene Technologies and its use is limited solely for research purposes.

Alternate Names for DGK-beta Overexpression Lysate

  • 90 kDa diacylglycerol kinase
  • DAG kinase beta
  • DAGK2
  • DGKB
  • DGKbeta
  • DGK-beta
  • diacylglycerol kinase beta
  • diacylglycerol kinase, beta (90kD)
  • diacylglycerol kinase, beta 90kDa
  • Diglyceride kinase beta
  • EC 2.7.1
  • EC 2.7.1.107
  • KIAA0718DGKDGK-BETA

Background

Diacylglycerol kinases (DGKs) are regulators of the intracellular concentration of the second messenger diacylglycerol (DAG) and thus play a key role in cellular processes. Nine mammalian isotypes have been identified, which are encoded by separate genes. Mammalian DGK isozymes contain a conserved catalytic (kinase) domain and a cysteine-rich domain (CRD). The protein encoded by this gene is a diacylglycerol kinase, beta isotype. Two alternatively spliced transcript variants have been found for this gene. (provided by RefSeq)

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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Bioinformatics

Gene Symbol DGKB