JMJD2D Overexpression Lysate

Western Blot: JMJD2D Overexpression Lysate (Adult Normal) [NBL1-12105] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for JMJD2D.

• Reactivity: Hu
• Applications: WB

JMJD2D Overexpression Lysate Summary

• Description:

  JMJD2D Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC212600
  
  Accession#: NM_018039
  
  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: KDM4D

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: 58.4 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.

Alternate Names for JMJD2D Overexpression Lysate

• EC 1.14.11
• EC 1.14.11.-
• FLJ10251
• JHDM3D
• JmjC domain-containing histone demethylation protein 3D
• JMJD2D
• JMJD2Dlysine-specific demethylase 4D
• jumonji domain containing 2D
• Jumonji domain-containing protein 2D
• KDM4D
• lysine (K)-specific demethylase 4D
• MGC141909

Background

JMJD2 is a member of the family of novel JmjC domain-containing histone demethylation (JHDM) enzymes, which has been shown to perform the enzymatic removal of methyl groups from histones. Histone demethylation by JHDM proteins requires cofactors Fe(II) and alpha-ketoglutarate. Family members include JHDM1 (demethylating histone 3 at lysine 36), and JHDM2A as well as JMJD2CH3K9 (both of which demethylate histone 3 at lysine 9). Histone demethylase activity has been linked to tumor development, decreases in cell proliferation, and hormone-dependent transcriptional activation.

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.

Bioinformatics

• Gene Symbol: KDM4D