Human LAP3 knockout A549 cell line
- Advanced Validation
- What is this?
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LAP3 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp insertion in exon 6.
View Alternative Names
AMPL_HUMAN, Cytosol aminopeptidase, HEL-S-106, LAP, LAPEP, Leucine aminopeptidase 3, Leucyl aminopeptidase, PEPS, Peptidase S, Proline aminopeptidase, Prolyl aminopeptidase, epididymis secretory protein Li 106
- WB
Lab
Western blot - Human LAP3 knockout A549 cell line (AB266986)
Lanes 1-4 : Merged signal (red and green). Green - ab154809 observed at 56 kDa. Red - loading control ab8245 observed at 36 kDa.
ab154809 Anti-LAP3 antibody [EPR10330] was shown to specifically react with LAP3 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab266986 (knockout cell lysate ab258023) was used. Wild-type and LAP3 knockout samples were subjected to SDS-PAGE. ab154809 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-LAP3 antibody [EPR10330] (<a href='/en-us/products/primary-antibodies/lap3-antibody-epr10330-ab154809'>ab154809</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
LAP3 knockout A549 cell lysate at 20 µg
Lane 2:
Western blot - Human LAP3 knockout A549 cell line (ab266986)
Lane 3:
HepG2 cell lysate at 20 µg
Lane 4:
HeLa 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: 56 kDa
Observed band size: 56 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human LAP3 knockout A549 cell line (AB266986)
Homozygous : 2 bp insertion in exon6
Reactivity data
Product details
Recommended control: Human wild-type A549 cell line (ab255450). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
Culture medium
F-12K + 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
LAP3 participates in protein maturation and degradation. It plays a role in processing peptides that are destined for further degradation or cellular signaling. The LAP3 protein is not known to be part of a larger complex but it works alongside other enzymes involved in peptide metabolism. This activity contributes to the regulation of intracellular peptide concentrations and supports cellular homeostasis.
Pathways
LAP3 is linked to important biological processes such as protein catabolism and the regulation of blood pressure. It interacts within the peptide processing pathways to influence the generation of bioactive peptides. LAP3 works in conjunction with other proteolytic enzymes such as aminopeptidases and endopeptidases which are essential for various cellular functions. Through these pathways LAP3 indirectly impacts the regulation of physiological processes.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB249116
Anti-LAP3 antibody [EPR10330] - BSA and Azide free
RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-LAP3 antibody [EPR10330] - BSA and Azide free (AB249116)](https://content.abcam.com/products/images/lap3-antibody-epr10330-bsa-and-azide-free-ab249116--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img90620.jpg)
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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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