Kelch-Like 3 Overexpression Lysate

Western Blot: Kelch-Like 3 Overexpression Lysate (Adult Normal) [NBP2-06980] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Kelch-Like 3.

• Reactivity: Hu
• Applications: WB

Kelch-Like 3 Overexpression Lysate Summary

• Description:

  Kelch-Like 3 Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC218393
  
  Accession#: NM_017415
  
  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: KLHL3

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: 64.8 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 Kelch-Like 3 Overexpression Lysate

• FLJ40871
• kelch-like 3 (Drosophila)
• kelch-like protein 3
• KIAA1129kelch (Drosophila)-like 3
• MGC44594

Background

KLHL3, also known as Kelch-like protein 3; has 3 isoforms, a 65 kDa 587 amino acid isoform A, a 61 kDa 555 amino acid isoform B, and a 56 kDa 505 amino acid isoform C; acts as a substrate-specific adapter of a BCR (BTB-CUL3-RBX1) E3 ubiquitin ligase complex that plays a role of regulator in ion transport of the distal nephron. The BCR(KLHL3) complex may regulate the SLC12A3/NCC subcellular location at the cell membrane by mediating ubiquitination of SLC12A3/NCC. There has been research revolving around the KLHL3 protein involvement in pharyngitis and pseudohypoaldosteronism. The protein has been shown to interact with OPRD1, GNAO1, SYT2, SYNCRIP, and GNAI1 proteins in protein ubiquitination, renal sodium ion absorption, and distal tubule morphogenesis pathways.

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: KLHL3