HOXB13 Overexpression Lysate

Western Blot: HOXB13 Overexpression Lysate (Adult Normal) [NBP2-07632] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for HOXB13.

• Reactivity: Hu
• Applications: WB

HOXB13 Overexpression Lysate Summary

• Description:

  HOXB13 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC209991
  
  Accession#: NM_006361
  
  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: HOXB13

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: 30.5 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 HOXB13 Overexpression Lysate

• homeo box B13
• homeobox B13
• homeobox protein Hox-B13
• HOXB13
• PSGD

Background

HOXB13 is a novel member of the AbdB subfamily of vertebrate HOX genes which are clustered in 4 unlinked complexes in the genome (HOXA, HOXB, HOXC, and HOXD clusters) which each span about 200 kb and contains 9 to 11 genes, transcribed from the same strand of DNA. The order of the HOX genes along the chromosome correlates with their expression along the anterior/posterior axis of the embryo and suggests that their organization is integral to their proper regulation. Members of the Abdominal B (AbdB) subfamily of homeobox genes exhibit posterior domains of expression, including the developing urogenital system, in vertebrates. Several HOXB genes have been described in humans and other mammals though it was thought that HOXB genes 10 through 13 had been lost in evolution until HOXB13 was discovered. HOXB13 gene contains 2 exons and encodes a 284-amino acid protein, 2 residues shorter than the mouse protein. It is separated from HOXB9 by 70 kb but is transcribed in the same orientation as the other HOXB genes. The 60-amino acid homeodomain, located near the C terminus of the protein, demonstrated 78 to 83% identity with HOX proteins in paralog group 13 of the AbdB subfamily; 6 of the amino acid differences from other member of this group represented conservative changes. HOXB13 had less than 60% identity to other AbdB-related genes. Mouse Hoxb13 is expressed first in the tailbud of embryonic mice and subsequently in the hindgut, urogenital tract, and the posterior extent of the spinal cord. It is not expressed in the secondary axes. During the first 2 trimesters of development, wound healing occurs without scars. Two homeobox genes, PRX2 and HOXB13, are differentially expressed during fetal versus adult wound healing. Both genes are expressed in proliferating fibroblasts and fetal dermis, but not in adult dermis. The HOXB13 gene has been mapped to human chromosome 17q21.2, where other HOXB genes are clustered, whilst the mouse Hoxb13 gene mapped to chromosome 11.

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: HOXB13