Human ERGIC3 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ERGIC3 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 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
2310015B14Rik, AV318804, C20orf47, CGI 54, D2Ucla1, DKFZp547A2190, ERGI3_HUMAN, ERGIC and golgi 3, ERV46, Endoplasmic reticulum-Golgi intermediate compartment protein 3, NY BR 84, PRO0989, RP23-220D12.2, RP3-477O4.1, SDBCAG84, Serologically defined breast cancer antigen 84, Serologically defined breast cancer antigen NY-BR-84, dJ477O4.2, endoplasmic reticulum localized protein ERp43
- WB
Lab
Western blot - Human ERGIC3 knockout HEK-293T cell line (AB266489)
Lanes 1-4 : Merged signal (red and green). Green - ab129179 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.
ab129179 Anti-ERGIC3 antibody [EPR8141] was shown to specifically react with ERGIC3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266489 (knockout cell lysate ab257941) was used. Wild-type and ERGIC3 knockout samples were subjected to SDS-PAGE. ab129179 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 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.
All lanes:
Western blot - Anti-ERGIC3 antibody [EPR8141] (<a href='/en-us/products/primary-antibodies/ergic3-antibody-epr8141-ab129179'>ab129179</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ERGIC3 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ERGIC3 knockout HEK-293T cell line (ab266489)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
MCF-7 cell lysate at 20 µg
Secondary
All lanes:
Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 50 kDa
false
- Cell Culture
Unknown
Cell Culture - Human ERGIC3 knockout HEK-293T cell line (AB266489)
Representative images of ERGIC3 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human ERGIC3 knockout HEK-293T cell line (AB266489)
Homozygous : 1 bp insertion 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, 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
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
The ERGIC3 protein functions as part of the larger ER-Golgi intermediate compartment (ERGIC) complex helping in the sorting and transport of proteins. The ERGIC serves as an essential hub in the secretory pathway modulating cargo movement to the Golgi for further processing. Some studies have implicated ERGIC3 in maintaining Golgi integrity and optimal function affecting overall cellular health.
Pathways
ERGIC3 plays a significant role in the secretory pathway assisting in the transit of proteins and the correct folding of polypeptides. It links functionally to other key proteins such as KI67 which associates with cell proliferation regions and relies heavily on efficient protein processing for cellular division. The association suggests ERGIC3's involvement in maintaining cellular growth and regeneration.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERGIC3 antibody [EPR8141] (AB129179)](https://content.abcam.com/products/images/ergic3-antibody-epr8141-ab129179--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img131314.jpg)
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