Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line
- Advanced Validation
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SIGMAR1 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 1. 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
AL024364, Aging-associated gene 8 protein, RP23 167I12.6, SGMR1_HUMAN, SIG-1R, SR-BP, SR31747-binding protein, Sigma 1-type opioid receptor, Sigma non-opioid intracellular receptor 1, Sigma1-receptor, Sigma1R, Sigmar1, hSigmaR1, mSigmaR1, opioid receptor, sigma 1, opioid receptor, sigma 1 isoform 1
- WB
Collaborator
Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line (AB266619)
ab253192 was shown to react with SIGMAR1 in wild-type HEK293T in Western blot with loss of signal observed in SIGMAR1 knockout cell line ab266619 (SIGMAR1 knockout cell lysate ab258666). Wild-type HEK293T and HeLa cell lysates and the corresponding SIGMAR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab253192 overnight at 4 ° at a 1/5000 dilution. Blots were incubated with goat anti-rabbit HRP secondary antibodies at 0.2μg/mL before imaging.
These data were provided by YCharOS Inc. an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.
All lanes:
Western blot - Anti-Sigma1-receptor antibody [EPR23266-69] (<a href='/en-us/products/primary-antibodies/sigma1-receptor-antibody-epr23266-69-ab253192'>ab253192</a>) at 1/5000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell lysate (<a href='/en-us/products/cell-lysates/human-sigmar1-sigma1-receptor-knockout-hek-293t-cell-lysate-ab258666'>ab258666</a>) at 20 µg
Lane 2:
Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line (ab266619)
Lane 3:
Wild-type HeLa cell lysate at 20 µg
Lane 4:
SIGMAR1 knockout HeLa cell lysate at 20 µg
Secondary
All lanes:
goat anti-rabbit HRP at 0.2 µg/mL
Predicted band size: 25 kDa
false
- WB
Lab
Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line (AB266619)
Lanes 1 - 4 : Merged signal (red and green). Green - ab151288 observed at 24 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab151288 was shown to react with Sigma1-receptor in wild-type HEK-293T cells in Western blot with loss of signal observed in SIGMAR1 knockout cell line ab266619 (SIGMAR1 knockout cell lysate ab258666). Wild-type HEK-293T and SIGMAR1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab151288 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ° at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
Lanes 1 - 4:
Western blot - Anti-Sigma1-receptor antibody (<a href='/en-us/products/primary-antibodies/sigma1-receptor-antibody-ab151288'>ab151288</a>) at 1/500 dilution
Lanes 1 - 4:
Western blot - Anti-Sigma1-receptor antibody (<a href='/en-us/products/primary-antibodies/sigma1-receptor-antibody-ab223702'>ab223702</a>) at 0.04 µg/mL
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
SIGMAR1 knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line (ab266619)
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
Caco-2 cell lysate at 20 µg
Predicted band size: 25 kDa
Observed band size: 24 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human SIGMAR1 (Sigma1-receptor) knockout HEK-293T cell line (AB266619)
Homozygous : 4 bp deletion in exon 1
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.
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
AB253192
Anti-Sigma1-receptor antibody [EPR23266-69]
BOND RX™ Validated|20ul selling size|RabMAb|Recombinant|KO Validated|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sigma1-receptor antibody [EPR23266-69] (AB253192)](https://content.abcam.com/products/images/sigma1-receptor-antibody-epr23266-69-ab253192--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img56839.jpg)
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- 0
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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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