Klotho Overexpression Lysate

Novus Biologicals | Catalog # NBL1-12307

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for Klotho Overexpression Lysate

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.

Product Specifications

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.

Type

Overexpression

Scientific Data Images for Klotho Overexpression Lysate

Western Blot: Klotho Overexpression Lysate [NBL1-12307]

Western Blot: Klotho Overexpression Lysate [NBL1-12307]

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.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: Klotho

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.

Alternate Names

KL

Gene Symbol

KL

Additional Klotho Products

Product Documents for Klotho Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Klotho Overexpression Lysate

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.

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.

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FAQs for Klotho Overexpression Lysate

Showing  1 - 2 的 2 FAQs Showing All

    • A: 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?

    • Q: 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.
      A: 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.
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