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Creative Biolabs

Human CTNNB1 Knockout HCT116 Cell Lysate

[CAT#: NCL2008ZP311] Review(5) Q&As(3)

Human CTNNB1 knockout cell lysate

Species:
Human
Cell Types:
Other Cells

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Product Overview

Description

This is a HCT116 cell transfected lysate in which Human CTNNB1 has been knockout by using CRISPR/Cas9 gene editing technology.

Cell Types

Other Cells

Cell Location

Epithelial

Application Notes

To use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.

*Usage of SDS sample buffer is not recommended with these lyophilized lysates.

Mutation Description

Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon3.

Passage Number

<20

Knockout Validation

Sanger Sequencing

Research Areas

Neurogenesis

Species

Human
Properties

Cell State

Cell lysate

STR Analysis

Amelogenin X
D5S818: 10, 11
D13S317: 10, 12
D7S820: 11, 12
D16S539: 11, 13
vWA: 17, 22
TH01: 8,9
TPOX: 8, 9
CSF1PO: 7, 10

Cell Purity

>95%

Storage

Store at -80°C. Please refer to protocols.

Research Use Only

For research use only, not for diagnostic or therapeutic use.
Target Details

Target

CTNNB1

Official Name

CTNNB1

Full Name

Catenin beta-1

Alternative Names

CTNNB1; CTNNB; MRD19; armadillo; catenin beta 1; EVR7; NEDSDV

Gene ID

1499 (Human); 12387 (Mouse); 84353 (Rat)

Uniprot ID

P35222 (Human); Q02248 (Mouse)
Publications

Publications (0)

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Customer Reviews and Q&As
Customer Reviews Average Customer Ratings Overall
5.0
user
Excellent
The lysate was easy to use, and the knockout was confirmed via Western blot
We observed significant changes in the expression of downstream targets, which provided clear insights into our experimental hypothesis. The consistency between batches was also commendable.
user
Excellent
We were able to reproduce our results consistently
It has saved us a considerable amount of time and effort that would otherwise have been spent creating our own knockout lines. The quality of the lysate is excellent, with minimal degradation and high protein content.
user
Excellent
Highly satisfied with this purchase
This lysate provided a reliable source of CTNNB1 knockout protein for our assays. The data obtained from using this product were robust and reproducible. We particularly appreciated the detailed datasheet that accompanied the product, which helped us integrate it seamlessly into our experimental workflows.
user
Excellent
We will certainly be using this product again in future experiments
It was essential for our comparative analyses between wild-type and knockout cell lines. The product's quality ensured that our Western blot results were clear and interpretable, leading to meaningful conclusions in our research.
user
Excellent
We utilized this lysate in our project
The knockout validation was spot on, and the lysate worked perfectly in our immunoprecipitation and Western blot experiments. The lysate's high quality contributed significantly to the reliability of our data.
Q&As
What is the origin and characterization of the lysate?
The cell lysate is derived from HCT116 cells, a human colorectal carcinoma cell line, which has been genetically modified to knock out the CTNNB1 gene. The knockout was achieved using CRISPR-Cas9 technology, ensuring a complete loss of β-catenin protein expression. The lysate is characterized by Western blotting to confirm the absence of β-catenin, and other relevant assays to ensure the integrity and utility of the lysate for neuroscience research.
How should I store the lysate to maintain its stability?
The cell lysate should be stored at -80°C immediately upon receipt to maintain its stability. Avoid repeated freeze-thaw cycles, as they can degrade the proteins and affect the lysate’s quality. If aliquoting, ensure that you use sterile, low-protein-binding tubes and work quickly to minimize exposure to room temperature.
Is there any specific protocol for using the cell lysate in Western blotting?
For WB, we recommend thawing the lysate on ice and preparing it with a suitable loading buffer. Typically, a protein concentration of 20-30 µg per lane is sufficient, but this can be optimized based on your specific requirements. Use appropriate primary and secondary antibodies for detecting β-catenin and other target proteins. Follow standard Western blotting procedures, ensuring all steps are performed at recommended conditions to achieve optimal results.
For Research Use Only. Not For Clinical Use.
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