Human AGO3 knockout HeLa cell line
- Advanced Validation
- What is this?
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AGO3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 12. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
5730550L01Rik, AGO3_HUMAN, Argonaute 3, EIF2C3 protein, Eukaryotic translation initiation factor 2C 3, FLJ12765, MGC86946, Protein argonaute-3, eIF-2C 3, hAgo3
- WB
Lab
Western blot - Human AGO3 knockout HeLa cell line (AB265320)
ab154844 Anti-AGO3 antibody [EPR9576] was shown to specifically react with Argonaute RISC Catalytic Component 3 in HeLa wild-type cells (ab255928). Loss of signal was observed when knockout cell line ab265320 (knockout cell lysate ab257819) was used. Wild-type and Argonaute RISC Catalytic Component 3 knockout samples were subjected to SDS-PAGE. ab154844 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Lanes 1 - 4:
Western blot - Anti-AGO3 antibody [EPR9576] (<a href='/en-us/products/primary-antibodies/ago3-antibody-epr9576-ab154844'>ab154844</a>) at 1/2000 dilution
Lane 3:
154844 at 1/2000 dilution
Lane 1:
Wild-type HeLa lysate at 40 µg
Lane 2:
Argonaute RISC Catalytic Component 3 knockout HeLa lysate at 40 µg
Lane 2:
Western blot - Human AGO3 knockout HeLa cell line (ab265320)
Lane 3:
Daudi lysate at 40 µg
Lane 4:
A549 lysate at 40 µg
Predicted band size: 97 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human AGO3 knockout HeLa cell line (AB265320)
Homozygous : 14 bp deletion in exon 12.
Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Zygosity
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
AGO3 contributes to gene expression regulation by silencing specific RNA transcripts. This protein forms a complex with other proteins and RNA molecules to execute its functions. Part of the miRNA and RNA interference pathways AGO3 associates with Dicer and other Argonaute proteins creating an effective silencing machinery that modulates various cellular processes. Its activity is essential for maintaining cellular homeostasis through RNA interference-mediated gene silencing.
Pathways
AGO3 plays a significant role in the RNA interference (RNAi) and the microRNA (miRNA) pathways. These pathways are key in post-transcriptional gene regulation controlling gene expression patterns important for development and differentiation. AGO3 interacts with Dicer a pivotal enzyme in the RNAi pathway facilitating the processing of precursor miRNAs into mature miRNAs further guiding RISC to its target mRNA sequences. These interactions highlight AGO3's critical place in RNA silencing networks.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Western blot - Anti-AGO3 antibody [EPR9576] (AB154844)](https://content.abcam.com/products/images/ago3-antibody-epr9576-ab154844--western-blot-img143878.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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