Human GOLM1 (GOLPH2) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human GOLM1 (GOLPH2) knockout HEK-293T cell line (AB266505)
False colour image of Western blot : Anti-GOLPH2 antibody [OTI6C9] staining at 1/500 dilution shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution shown in red. In Western blot ab119800 was shown to bind specifically to GOLPH2. A band was observed at 70-75 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in GOLM1 knockout cell line ab266505 (knockout cell lysate ab257976). To generate this image wild-type and GOLM1 knockout HEK-293T cell lysates were analysed. First 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°. 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-GOLPH2 antibody [OTI6C9] (<a href='/en-us/products/primary-antibodies/golph2-antibody-oti6c9-ab119800'>ab119800</a>) at 1/500 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 10 µg
Lane 2:
GOLM1 knockout HEK-293T cell lysate at 10 µg
Lane 2:
Western blot - Human GOLM1 (GOLPH2) knockout HEK-293T cell line (ab266505)
Lane 3:
U-2 OS cell lysate at 20 µg
Lane 4:
HepG2 cell lysate at 20 µg
Lane 5:
MDA-MB-231 cell lysate at 20 µg
Lane 6:
LNCaP cell lysate at 20 µg
Predicted band size: 45 kDa
Observed band size: 70-75 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human GOLM1 (GOLPH2) knockout HEK-293T cell line (AB266505)
Homozygous : 1 bp insertion in exon 3
- Cell Culture
Unknown
Cell Culture - Human GOLM1 (GOLPH2) knockout HEK-293T cell line (AB266505)
Representative images of GOLM1 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.
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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
GOLPH2 plays an essential role in maintaining Golgi function and efficient protein trafficking. It does not belong to a large protein complex but its proper functioning is critical for handling secretory pathways. The protein keeps secretory flow organized which affects cellular responses and interactions with the environment. The GOLPH2 protein also has connections to immunity responding to stimuli like infections or inflammation.
Pathways
GOLPH2 actively participates in secretory pathways and protein transport systems. Its role includes coordination with other Golgi proteins to facilitate protein modification and sorting. GOLPH2 interacts with other proteins like GOLM1 and GOLPH3 impacting the Golgi's structural organization and influencing pathways like vesicular transport. These interactions are important for adapting the intracellular environment to external demands.
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'.
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