HACE1 Overexpression Lysate

Western Blot: HACE1 Overexpression Lysate (Adult Normal) [NBL1-11433] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for HACE1.

• Reactivity: Hu
• Applications: WB

HACE1 Overexpression Lysate Summary

• Description:

  HACE1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC205602
  
  Accession#: NM_020771
  
  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: HACE1

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: 102.2 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 HACE1 Overexpression Lysate

• E3 ubiquitin-protein ligase HACE1
• EC 6.3.2
• HACE
• HECT domain and ankyrin repeat containing, E3 ubiquitin protein ligase 1
• HECT domain and ankyrin repeat-containing E3 ubiquitin-protein ligase 1
• KIAA1320EC 6.3.2.-

Background

The HACE1 gene codes for a E3 ubiquitin-protein ligase HACE1 that is a participant in Golgi membrane fusion as well as in regulation of small GTPases to mediate ubiquitination and degradation of active RAC1, working in defense against pathogens. This protein may also play a role as a transcription regulator through its interaction with RARB. HACE1 exists in four isoforms: isoform 1 is 909 amino acids long, 102 kDA; isoform 2 is 318 amino acids long, 35 kDA; isoform 3 is 562 amino acids long, 63 kDA; isoform 4 is 694 amino acids long, nearly 78 kDA. HACE1 has been researched in regards to celiac disease, malaria, colorectal cancer, wilms tumors, and carcinoma. It is involved in the SMAD signaling network as well as ubiquitin-proteasome dependent proteolysis and has been known to interact with genes RARA, RAB4A, RAC1, PLXNA2, and IRF7.

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: HACE1