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DENND1B Overexpression Lysate

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Western Blot: DENND1B Overexpression Lysate (Adult Normal) [NBL1-08314] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DENND1B.
Western Blot: DENND1B Overexpression Lysate (Adult Normal) [NBL1-08314] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for DENND1B.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications WB

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DENND1B Overexpression Lysate Summary

Description

DENND1B Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC210363

Accession#: NM_019049

Protein Tag: C-MYC/DDK

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
DENND1B

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
37.2 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.

Alternate Names for DENND1B Overexpression Lysate

  • C1ORF18
  • C1orf218
  • chromosome 1 open reading frame 218
  • Connecdenn 2
  • DENN domain-containing protein 1B
  • DENN/MADD domain containing 1B
  • DKFZp547O0715
  • FAM31BMGC103810
  • FLJ20054
  • FLJ21179
  • FLJ33888
  • member B
  • Protein FAM31B

Background

DENND1B is a member of the connecdenn family, which links RAB35 activation with the clathrin machinery. The protein functions as a guanine nucleotide exchange factor for RAB35. DENND1B is present at higher levels in dendritic and natural killer cells but at lower levels in the other myeloid lineage cells.

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 DENND1B