KLF15 Overexpression Lysate

Western Blot: KLF15 Overexpression Lysate (Adult Normal) [NBL1-12317] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for KLF15.

• Reactivity: Hu
• Applications: WB

KLF15 Overexpression Lysate Summary

• Description:

  KLF15 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC207523
  
  Accession#: NM_014079
  
  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: KLF15

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: 43.8 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 KLF15 Overexpression Lysate

• DKFZp779M1320
• kidney-enriched Kruppel-like factor
• KKLFKidney-enriched krueppel-like factor
• Krueppel-like factor 15
• Kruppel-like factor 15

Background

The kidney chloride channel proteins CLCNKA and CLCNKB are highly related, although they are located in different areas of the kidney. Using a yeast 1-hybrid screen of a kidney cDNA library with the GA element of the CLCNKA promoter as bait, Uchida et al. (2000) isolated cDNAs encoding MAZ and KLF15, which they termed KKLF. Sequence analysis predicted that the 415-amino acid KLF15 protein, which is 84% identical to the rat Klf15 protein, contains 3 zinc finger motifs at its C terminus, N-terminal serine-rich stretches, and a central proline-rich segment. EMSA analysis confirmed that KLF15 and MAZ interact with the GA element of the CLCNKA promoter and showed that KLF15 binds with higher affinity and is a functional competitor of MAZ. Northern blot analysis revealed highest expression of a 2.5-kb KLF15 transcript in liver, followed by heart, skeletal muscle, and kidney. No expression was found in bone marrow or lymphoid tissues. Western blot analysis showed expression of a 50-kD protein. Immunohistochemical analysis detected nuclear expression of KLF15 in liver sinusoid stellate cells, subcapsular fibroblasts, and portal fibroblasts. KLF15 was also expressed in cardiac and skeletal muscle interstitial cells and in kidney inner medulla, glomeruli, and cortical interstitium. Immunofluorescence microscopy, however, demonstrated no colocalization of KLF15 with CLCNKA.

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: KLF15