Human RCN1 (RCN) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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RCN1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 1.
View Alternative Names
Epididymis secretory protein Li 84, FLJ37041, HEL S 84, PIG20, Proliferation inducing gene 20, RCAL, RCN, RCN1_HUMAN, Reticulocalbin 1 EF hand calcium binding domain, Reticulocalbin-1
- WB
Lab
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (AB266098)
Lanes 1- 4 : Merged signal (red and green). Green - ab210404 observed at 45 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab210404 was shown to react with RCN1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266098 (knockout cell lysate ab258172) was used. Wild-type HEK-293T and RCN1 knockout HEK-293T 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. ab210404 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-RCN1/RCN antibody [EPR17163-117] (<a href='/en-us/products/primary-antibodies/rcn1-rcn-antibody-epr17163-117-ab210404'>ab210404</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
RCN1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (ab266098)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Daudi cell lysate at 20 µg
Predicted band size: 39 kDa
Observed band size: 45 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RCN1 (RCN) knockout HEK-293T cell line (AB266098)
Homozygous : 1 bp insertion in exon 1
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
RCN1 plays a role in cellular calcium homeostasis and may influence processes such as protein folding and trafficking. It is not known to be a part of a larger complex but it interacts with other ER-resident proteins. Maintaining calcium levels is important within the ER where RCN1 helps buffer calcium concentrations. This function supports processes that are sensitive to calcium fluctuations such as signal transduction and stress responses.
Pathways
The role of RCN1 in cellular mechanisms connects it to important pathways like the calcium signaling pathway and the unfolded protein response pathway. In the calcium signaling pathway RCN1 aids in regulating calcium levels within the ER affecting downstream signaling events. Meanwhile in the unfolded protein response pathway RCN1 participates by ensuring the proper folding of proteins particularly during times of stress when misfolded proteins accumulate. RCN1's interaction with calumenin another calcium-binding protein highlights its involvement in these pathways.
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
AB210404
Anti-RCN1/RCN antibody [EPR17163-117]
RabMAb|Recombinant|KO Validated
![Western blot - Anti-RCN1/RCN antibody [EPR17163-117] (AB210404)](https://content.abcam.com/products/images/rcn1-rcn-antibody-epr17163-117-ab210404--western-blot-img7213.jpg)
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Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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