HMGB1/HMG-1 Overexpression Lysate

Western Blot: HMGB1 Overexpression Lysate (Adult Normal) [NBL1-11614] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for HMGB1.

• Reactivity: Hu
• Applications: WB

HMGB1/HMG-1 Overexpression Lysate Summary

• Description:

  HMGB1/HMG-1 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC205918
  
  Accession#: NM_002128
  
  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: HMGB1

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: 24.7 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 HMGB1/HMG-1 Overexpression Lysate

• Amphoterin
• high mobility group box 1
• High mobility group protein 1
• high mobility group protein B1
• high-mobility group (nonhistone chromosomal) protein 1
• high-mobility group box 1
• HMG1
• HMG-1
• HMG1DKFZp686A04236
• HMG3
• HMGB1
• SBP-1
• Sulfoglucuronyl carbohydrate binding protein

Background

HMGB1 and HMGB2 are part of the chromatin non-histone high mobility group proteins 1 and 2. HMGB1 and HMGB2 (containing multiple HMG-boxes) are conserved domains of 80 amino acids which mediate the DNA binding of many proteins. HMG box domains recognize DNA structure. Both HMGB1 and HMGB2 contain a N-terminal HMG box, a central HMG box, and an acidic carboxy terminus. The acidic tails of HMGB1 and HMGB2 contain multiple serine residues which match the phosphorylation consensus sites of casein kinase II, and phosphorylation of this domain appears to be important for proper functioning of these proteins. HMGB1 and HMGB2 have been shown to facilitate the binding of various sequence-specific transcription factors to their respective DNA binding sites. HMGB1 and HMGB2 may also serve as architectural factors that recognize and mediate DNA structural changes that accompany various events such as DNA repair, transcription, and replication.

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: HMGB1