Klotho Overexpression Lysate

Western Blot: Klotho Overexpression Lysate (Adult Normal) [NBL1-12307] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for Klotho.

• Reactivity: Hu
• Applications: WB

Klotho Overexpression Lysate Summary

• Description:

  Klotho Transient Overexpression Lysate

  
  Expression Host: HEK293T
  
  Plasmid: RC224132
  
  Accession#: NM_004795
  
  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: KL

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: 116 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 Klotho Overexpression Lysate

• EC 3.2.1
• EC 3.2.1.31
• KL
• Klotho

Background

This gene encodes a type-I membrane protein that is related to beta-glucosidases. Reduced production of this protein has been observed in patients with chronic renal failure (CRF), and this may be one of the factors underlying the degenerative processes (e.g., arteriosclerosis, osteoporosis, and skin atrophy) seen in CRF. Also, mutations within this protein have been associated with ageing and bone loss. [provided by RefSeq]. Sequence Note: The RefSeq transcript and protein were derived from transcript and genomic sequence to make the sequence consistent with the reference genome assembly. The genomic coordinates used for the transcript record were based on alignments. Publication Note: This RefSeq record includes a subset of the publications that are available for this gene. Please see the Entrez Gene record to access additional publications.

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.

FAQs for Klotho Lysate (NBL1-12307). (Showing 1 - 2 of 2 FAQ).

1. I study physiology (PhD) and I need a klotho kit.
   • A list of our klotho products can be found here. I am not entirely sure what you are looking for as I don't have your complete message for some reason. Can you please clarify what you are looking for?
2. What antibody was used to validate this lysate (NBL1-12307) as a positive control? Klotho should run at approximately 116 kDa (as in this blot), yet none of the antibodies you sell show a band at the correct size. I want the antibody used to make this blot.
   • We tested our Klotho overexpression lysate with an anti-DDK antibody to detect the DDK tag attached to the transfected protein. For our Klotho antibodies: NBP1-76511 detects a band at approximately 200kDa. Our lab notes state that due to the numerous glycosylation sites on the human klotho protein, the band appears higher than expected. NBP1-55097 recognized the 62kDa isoform 2 of the human klotho protein. NBP1-22497 is a monoclonal that we have licensed. We unfortunately do not have any images available at this time, but have validated the listed applications due to publications utilizing this same clone.

Bioinformatics

• Gene Symbol: KL