Knockout Cell Lysate
Product DetailsApplicationCase StudyFAQ
Product Details
Engineered using CRISPR technology,these cell lines are rigorously verified via PCR and sequencing to confirm the double-knockout genotype.The resulting knockout cell lysate,prepared in RIPA buffer,includes a parental cell line lysate for internal control.Primarily serving as negative controls for western blots,these lysates validate antibody specificity,crucial for eliminating nonspecific binding.With potential applications as negative controls for functional assays,Creative Biogene's Knockout Cell Lysate series offers researchers a reliable solution for
precise genotype confirmation and experimental validation.
Quality Advantages
- Precise Genotype Confirmation: Each cell line undergoes rigorous PCR and sequencing to verify
its double-knockout genotype, ensuring accuracy and reliability.
- Consistent Lysate Preparation: The KO cell lysate is uniformly prepared in RIPA buffer,
maintaining consistency across experiments and facilitating reliable results.
- Comprehensive Quality Control: The inclusion of lysate from the parental cell line as an
internal control ensures the integrity of experimental comparisons, enhancing confidence in study outcomes.
- Application Versatility: Our products serve as invaluable negative controls for western
blotting, enabling robust antibody specificity validation and minimizing nonspecific binding concerns.
- Potential for Novel Applications: Anticipated novel uses include employing our products as
negative controls for functional assays, and expanding their utility and relevance in diverse research contexts.
Knockout Cell Lysate List
Application
knockout cell lines and their lysates serve as invaluable tools for quality control in various experimental assays.
They provide researchers with reliable negative controls for western blotting, ensuring the specificity and accuracy
of antibody detection. Moreover, these products facilitate the validation of drug targets by enabling the assessment
of target protein function upon gene knockout. Furthermore, their application extends to biomarker discovery, where
they aid in the identification and validation of biomolecules with diagnostic or prognostic significance.
Specifically, knockout cell lines and their lysates are indispensable resources that contribute to the advancement
of scientific research across diverse fields:
- Validating Antibodies: KO cell lines and lysate present an ideal platform for validating both
polyclonal and monoclonal antibodies, ensuring accuracy and reliability in antibody-based assays.
- Analyzing Gene Function and Biological Pathways: Utilizing KO cell lines and lysate facilitates
in-depth investigations into gene function and the intricate biological pathways involved, offering insights
into cellular mechanisms and interactions.
- Discovering and Validating Biomarkers: KO cell lines and lysate streamline the process of
discovering and validating biomarkers, contributing to identifying novel diagnostic and prognostic indicators in
various biological contexts.
- Identifying and Validating Drug Targets: Leveraging KO cell lines and lysate aids in the
identification and validation of potential drug targets, crucial for advancing drug discovery efforts and
developing targeted therapeutic interventions.
- Investigating Cancer Development and Progression: KO cell lines and lysate are invaluable tools
for elucidating the underlying mechanisms driving cancer development and progression, paving the way for novel
insights into tumor biology and therapeutic strategies.
Case Study
Case Study 1
Despite growing evidence of the importance of NAD+ levels for retinal health, the specific roles of subcellular NAD+
pools in retinal development, maintenance, and disease are still unclear. Researchers investigated the role of
NMNAT1 in the mammalian retina and found that its deletion led to severe degeneration of photoreceptors and inner
retinal neurons through various cell death pathways. To understand the underlying mechanisms, they analyzed
disruptions in retinal central carbon metabolism, purine nucleotide synthesis, and amino acid pathways.
Transcriptomic and immunostaining approaches also revealed dysregulation of photoreceptor and synapse-specific genes
even before morphological or metabolic changes were detectable. These findings suggest that NMNAT1 plays a crucial
role in maintaining retinal health and function, possibly by regulating gene expression during photoreceptor
terminal differentiation.
Figure 1. Knockout Cell Lysate is utilized by researchers to evaluate
the impact of gene knockout on cellular processes. It allows for the assessment of protein expression levels, such
as NMNAT1, and enables the comparison of cellular morphology between knockout and control samples using techniques
like Western blotting and histological analysis. (Sokolov D, et al., 2021)
Case Study 2
Prion diseases are deadly neurodegenerative disorders affecting both humans and animals, which can spread within and
occasionally across species. Researchers investigated the impact of autophagy modulation on the release of prions
packaged in exosomes and its implications for prion infection. Prions, known for causing fatal neurodegenerative
diseases, can spread via exosomes present in bodily fluids. By studying murine neuronal cells, they found that
stimulating autophagy inhibited exosomal prion release, while inhibiting autophagy increased exosome-mediated prion
dissemination. Moreover, differences in basal autophagy levels between cell types influenced the amount of exosomal
prion release. These findings highlight a novel role of autophagy in the prion life cycle, suggesting that
manipulating autophagy could be a potential strategy for controlling prion diseases.
Figure 2. Researchers used CRISPR/Cas9 to create Atg5-deficient ScN2a
cells (KO) lacking autophagy. Despite normal viability, Atg5-KO cells exhibited significantly higher exosome release
compared to WT cells, with increased PrPSc in exosomes but unchanged levels in knockout cell lysates. This suggests
that autophagy blockage enhances prion spread via exosomes. (Abdulrahman BA, et al., 2018)
FAQ
Q: How does Creative Biogene ensure the quality and reliability of its Knockout Cell Lysate series?
A: We ensure the quality and reliability of our Knockout Cell Lysate series through several measures:
(1) Rigorous PCR and sequencing verification of the double-knockout genotype
for each cell line.
(2) Uniform preparation of lysates in RIPA buffer, maintaining consistency
across experiments.
(3) Inclusion of lysate from the parental cell line as an internal control,
ensuring integrity in experimental comparisons.
Q: What are the potential applications of Creative Biogene's Knockout Cell Lysate series beyond western blotting?
A:Beyond western blotting,our Knockout Cell Lysate series holds potential for novel applications,including:
(1)Serving as negative controls for functional assays,ensuring specificity and accuracy in various experimental setups.
(2)Facilitating the validation of drug targets by assessing target protein function upon gene knockout,aiding in drug discovery efforts.
(3)Contributing to biomarker discovery by identifying and validating biomolecules with diagnostic or prognostic significance.
Q:How can researchers utilize Creative Biogene's Knockout Cell Lysate series in antibody validation
studies?
A: Researchers can utilize our Knockout Cell Lysate series as an ideal platform for validating both
polyclonal and monoclonal antibodies. These lysates ensure accuracy and reliability in antibody-based
assays by providing reliable negative controls for specificity testing.
Q: In what ways can Creative Biogene's Knockout Cell Lysate series contribute to the study of cancer development and progression?
A:Our Knockout Cell Lysate series can significantly contribute to elucidating the underlying mechanisms driving cancer development and progression.By providing reliable tools for gene knockout studies,these lysates enable researchers to gain novel insights into tumor biology,paving the way for the development of innovative therapeutic strategies.
* For research use only. Not intended for any clinical use.