Human MMP3 knockout U-87 MG cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human MMP3 knockout U-87 MG cell line (AB306727)
Western blot : Anti-MMP3 antibody [EP1186Y] (ab52915) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab52915 was shown to bind specifically to MMP3. A band was observed at 55-60 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in MMP3 knockout cell line. To generate this image, wild-type and MMP3 knockout U-87 MG cell lysates were analysed. First, 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes:
Western blot - Anti-MMP3 antibody [EP1186Y] (<a href='/en-us/products/primary-antibodies/mmp3-antibody-ep1186y-ab52915'>ab52915</a>) at 1/1000 dilution
Lane 1:
Wild-type U-87 MG cell lysate at 20 µg
Lane 2:
MMP3 knockout U-87 MG cell lysate at 20 µg
Lane 3:
Wild-type HeLa Treated: IL-1beta (10 ng/mL, 48 h) cell lysate at 20 µg
Lane 4:
Wild-type HeLa Vehicle control: IL-1beta (0 ng/mL, 48 h) cell lysate at 20 µg
Lane 5:
HeLa cell lysate at 20 µg
Lane 6:
MCF7 cell lysate at 20 µg
Secondary
All lanes:
Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 53 kDa
Observed band size: 55 kDa,60 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human MMP3 knockout U-87 MG cell line (AB306727)
8 bp deletion after Leu78 of the WT protein
Reactivity data
Product details
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
EMEM + 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
MMP3 contributes to multiple physiological and pathological processes. It supports the remodeling of connective tissues and assists in normal developmental processes. MMP3 can work as part of a complex with other MMP proteins which can enhance or inhibit its activity. The balance of MMPs including MMP3 regulates the extracellular matrix turnover and is important for maintaining tissue homeostasis.
Pathways
MMP3 participates in pathways involved in tissue remodeling and inflammatory responses. It is part of cellular pathways regulated by cytokines like interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha). These pathways link MMP3 to other proteins such as MMP1 and MMP9 which perform overlapping and distinct roles in matrix degradation during inflammation and tissue repair.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
![Western blot - Anti-MMP3 antibody [EP1186Y] (AB52915)](https://content.abcam.com/products/images/mmp3-antibody-ep1186y-ab52915--western-blot-img433953.jpg)
Product promise
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