Human ACTN1 (alpha Actinin) knockout HeLa cell line
- Advanced Validation
- What is this?
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ACTN1 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: 4 bp deletion in exon 2. 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
ACTN1_HUMAN, Actinin alpha 1, Alpha-actinin cytoskeletal isoform, Alpha-actinin-1, BDPLT15, F-actin cross-linking protein, FLJ40884, FLJ54432, Non-muscle alpha-actinin-1, actinin 1 smooth muscle
- WB
Lab
Western blot - Human ACTN1 (alpha Actinin) knockout HeLa cell line (AB265610)
Lanes 1- 2 : Merged signal (red and green). Green - ab68194 observed at 103 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab68194 was shown to react with alpha Actinin/ACTN1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265610 (knockout cell lysate ab257337) was used. Wild-type HeLa and ACTN1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab68194 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 2000 dilution and a 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.
All lanes:
Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-actinin-actn1-antibody-ep2527y-loading-control-ab68194'>ab68194</a>) at 1/2000 dilution
Lane 1:
Wild-type HeLa cell lysate at 20 µg
Lane 2:
ACTN1 knockout HeLa cell lysate at 20 µg
Lane 2:
Western blot - Human ACTN1 (alpha Actinin) knockout HeLa cell line (ab265610)
Predicted band size: 103 kDa
Observed band size: 103 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ACTN1 (alpha Actinin) knockout HeLa cell line (AB265610)
Homozygous : 4 bp deletion in exon 2.
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
Alpha Actinin-1 helps maintain the structural integrity of cells by anchoring actin filaments and connecting them to integrins or membrane proteins. It often functions as part of a protein complex interacting with other cytoskeletal components and signaling molecules to stabilize cellular structures. In muscle cells it blends with titin and other sarcomeric proteins to assist muscle contraction. Beyond structural roles it partakes in cellular signaling pathways by interacting with signaling proteins and influencing cellular responses to external stimuli.
Pathways
Alpha Actinin-1 is involved in the regulation of the actin cytoskeleton pathway as well as the integrin signaling pathway. In these pathways it collaborates with proteins such as vinculin and paxillin modulating cell adhesion and movement. Its ability to interact with integrin-linked kinases suggests a role in transducing mechanical signals from the extracellular matrix to downstream signaling cascades.
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
Primary Antibodies
AB68194
Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - Loading Control
RabMAb|Recombinant|KO Validated|Lab Essentials
![Immunoprecipitation - Anti-alpha Actinin/ACTN1 antibody [EP2527Y] - Loading Control (AB68194)](https://content.abcam.com/products/images/alpha-actinin-actn1-antibody-ep2527y-loading-control-ab68194--immunoprecipitation-img11208.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com