KLHDC2 Overexpression Lysate

Western Blot: KLHDC2 Overexpression Lysate (Adult Normal) [NBL1-12325] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KLHDC2.

• Reactivity: Hu
• Applications: WB

KLHDC2 Overexpression Lysate Summary

• Description:

  KLHDC2 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC200116
  
  Accession#: NM_014315
  
  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: KLHDC2

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: 45.9 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 KLHDC2 Overexpression Lysate

• HCA33
• HCLP-1Host cell factor-like protein 1
• Hepatocellular carcinoma-associated antigen 33
• Host cell factor homolog LCP
• kelch domain containing 2
• kelch domain-containing protein 2
• KLHDC2
• LCP

Background

KLHDC2, also known as Kelch domain-containing protein 2, is a 46kDa 406 amino acid protein with a shorter 28kDa 249 isoform produced by alternative splicing. KLHDC2 interfers with CREB3-DNA binding by repressing CREB3-mediated transcription. Current research is being conducted on KLHDC2 in relation to Legg-Calve-Perthes disease, hepatocellular carcinoma and herpes simplex. KLHDC2 is linked to the reflex, excretion, micturition, cytokine production and lactation pathways where it interacts with DDX19B, SFN, STRN4, EZH2 and UBXN7.

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: KLHDC2