Human ERP29 knockout HEK-293T cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human ERP29 knockout HEK-293T cell line (AB266543)
Lanes 1-3 : Merged signal (red and green). Green - ab175193 observed at 29 kDa. Red - loading control ab8245 observed at 37 kDa.
ab175193 Anti-ERp29 antibody [EPR12985] was shown to specifically react with ERp29 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266543 (knockout cell lysate ab257188) was used. Wild-type and ERp29 knockout samples were subjected to SDS-PAGE. ab175193 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ERp29 antibody [EPR12985] (<a href='/en-us/products/primary-antibodies/erp29-antibody-epr12985-ab175193'>ab175193</a>) at 1/500 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ERP29 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ERP29 knockout HEK-293T cell line (ab266543)
Lane 3:
HepG2 cell lysate at 20 µg
Predicted band size: 29 kDa
Observed band size: 29 kDa
false
- WB
Lab
Western blot - Human ERP29 knockout HEK-293T cell line (AB266543)
Lanes 1-3 : Merged signal (red and green). Green - ab176573 observed at 29 kDa. Red - loading control ab8245 observed at 37 kDa.
ab176573 Anti-ERp29 antibody [EPR12499(B)] was shown to specifically react with ERp29 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266543 (knockout cell lysate ab257188) was used. Wild-type and ERp29 knockout samples were subjected to SDS-PAGE. ab176573 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 10000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-ERp29 antibody [EPR12499(B)] (<a href='/en-us/products/primary-antibodies/erp29-antibody-epr12499b-ab176573'>ab176573</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK293T cell lysate at 20 µg
Lane 2:
ERP29 knockout HEK293T cell lysate at 20 µg
Lane 2:
Western blot - Human ERP29 knockout HEK-293T cell line (ab266543)
Lane 3:
HepG2 cell lysate at 20 µg
Predicted band size: 29 kDa
Observed band size: 29 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human ERP29 knockout HEK-293T cell line (AB266543)
Homozygous : 1 bp insertion in exon 1
- Cell Culture
Unknown
Cell Culture - Human ERP29 knockout HEK-293T cell line (AB266543)
Representative images of ERP29 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 a EVOS XL Core microscope.
Complementary Products
Primary Antibodies
AB249928
Anti-ERp29 antibody [EPR12499(B)] - BSA and Azide free
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERp29 antibody [EPR12499(B)] - BSA and Azide free (AB249928)](https://content.abcam.com/products/images/erp29-antibody-epr12499b-bsa-and-azide-free-ab249928--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img422701.jpg)
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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
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
This protein assists in maintaining ER function by preventing the accumulation of misfolded proteins. ERp29 is part of the protein disulfide isomerase family which helps in forming disulfide bonds in proteins ensuring their correct structure and function. It does not form part of a larger complex but interacts with other proteins to manage stress and maintain cellular function. These interactions help in the secretory pathway important for processing proteins.
Pathways
ERp29 plays an innovative role in the Unfolded Protein Response (UPR) a cellular reaction to ER stress. It works alongside proteins like GRP78 within the UPR to mitigate stress's harmful effects by enhancing the protein folding capacity of the ER. Additionally ERp29's involvement in the protein disulfide isomerase family connects it to the oxidative protein folding pathway which is essential for maintaining redox balance in the ER.
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
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