Human RCN1 (RCN) knockout HEK-293T cell line
- Advanced Validation
- 각 태그의 뜻
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RCN1 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: 1 bp insertion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
대체 명칭 보기
RCN1, Reticulocalbin-1, RCN
- 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='/ko/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
- WB
Lab
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (AB266098)
Western blot : Anti-RCN1/RCN antibody [EPR17162] ab205927 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type HEK-293T cell lysates with no signal observed at this size in RCN1 knockout HEK-293T cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-RCN1/RCN antibody [EPR17162] (<a href='/ko/products/primary-antibodies/rcn1-rcn-antibody-epr17162-ab205927'>ab205927</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysates at 20 µg
Lane 2:
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (ab266098) at 20 µg
Lane 3:
HeLa cell lysates at 20 µg
Lane 4:
Daudi cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa
false
- WB
Lab
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (AB266098)
Western blot : Anti-RCN1/RCN antibody [EPR17163-117] ab210404 staining at 1/2000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type HEK-293T cell lysates with no signal observed at this size in RCN1 knockout HEK-293T cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-RCN1/RCN antibody [EPR17163-117] (<a href='/ko/products/primary-antibodies/rcn1-rcn-antibody-epr17163-117-ab210404'>ab210404</a>) at 1/2000 dilution
Lane 1:
Wild-type HEK-293T cell lysates at 20 µg
Lane 2:
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (ab266098) at 20 µg
Lane 3:
HeLa cell lysates at 20 µg
Lane 4:
Daudi cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa
false
- WB
Lab
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (AB266098)
Western blot : Anti-RCN1/RCN antibody [EPR17163] - C-terminal ab198996 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 45 kDa in Wild-type HEK-293T cell lysates with no signal observed at this size in RCN1 knockout HEK-293T cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-RCN1/RCN antibody [EPR17163] - C-terminal (<a href='/ko/products/primary-antibodies/rcn1-rcn-antibody-epr17163-c-terminal-ab198996'>ab198996</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysates at 20 µg
Lane 2:
Western blot - Human RCN1 (RCN) knockout HEK-293T cell line (ab266098) at 20 µg
Lane 3:
HeLa cell lysates at 20 µg
Lane 4:
Daudi cell lysates at 20 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa
false
보완 제품
Reactivity 정보
제품 세부 정보
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.
제품 구성
특성 및 보관 정보
유전자명
유전자 편집 종류
유전자 편집 방식
Knockout 검증
Zygosity
배송 시 보관 조건
적절한 단기 보관 조건
적절한 장기 보관 조건
추가 정보
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.
품질 관리
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
초기 취급 지침
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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>
Subculture 가이드라인
- 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.
배양 배지
DMEM (High Glucose) + 10% FBS
동결보존 배지
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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