Human LAMP1 knockout HCT116 cell line
- Advanced Validation
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LAMP1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided.
View Alternative Names
CD107 antigen-like family member A, CD107a, CD107a antigen, LAMP1_HUMAN, LAMPA, LGP120, Lysosomal membrane glycoprotein 120KD, Lysosomal Associated Membrane Protein 1, Lysosome-associated membrane glycoprotein 1, Lysosome-associated membrane protein 1, OTTHUMP00000040663, lgpA
- WB
Lab
Western blot - Human LAMP1 knockout HCT116 cell line (AB289151)
Western blot : Anti-LAMP1 antibody [EPR24395-31] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab278043 was shown to bind specifically to LAMP1. A band was observed at 100-120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout cell line. To generate this image, wild-type and LAMP1 knockout HCT 116 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-LAMP1 antibody [EPR24395-31] - Lysosome Marker (<a href='/en-us/products/primary-antibodies/lamp1-antibody-epr24395-31-lysosome-marker-ab278043'>ab278043</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
LAMP1 knockout HCT 116 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Observed band size: 100-120 kDa
false
- WB
Lab
Western blot - Human LAMP1 knockout HCT116 cell line (AB289151)
Western blot : Anti-LAMP1 antibody [EPR4204] - Lysosome Marker staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108597 was shown to bind specifically to LAMP1. A band was observed at 100-120 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in LAMP1 knockout cell line. To generate this image, wild-type and LAMP1 knockout HCT 116 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-LAMP1 antibody [EPR4204] - Lysosome Marker (<a href='/en-us/products/primary-antibodies/lamp1-antibody-epr4204-lysosome-marker-ab108597'>ab108597</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 cell lysate at 20 µg
Lane 2:
LAMP1 knockout HCT 116 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
MCF7 cell lysate at 20 µg
Observed band size: 100-120 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human LAMP1 knockout HCT116 cell line (AB289151)
55 bp deletion (allele 1) and 53 bp deletion (allele 2) after Cys40 of the WT protein
Reactivity data
Product details
Recommended control: Human wild-type HCT116 cell line (ab288559). 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
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
McCoY5a + 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
LAMP1 functions to protect lysosomal membranes from the harsh environment inside the lysosome. It forms part of a protective glycocalyx composed of highly glycosylated proteins supporting lysosomal stability. This protein interacts closely with other lysosome markers and assists in the fusion of vesicles with lysosomes making it fundamental to lysosomal processes. As a lysosome marker LAMP1 helps identify lysosomal compartments during studies involving microscopy and LAMP1 staining.
Pathways
The protein LAMP1 participates in the autophagy and endocytic pathways. It acts collaboratively within the lysosomal degradation route involving proteins like LAMP2 which also supports lysosomal function. LAMP1 influences these pathways by mediating the fusion of autophagosomes or endosomes with lysosomes thereby ensuring efficient breakdown of cellular debris and macromolecules.
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
AB24170
Anti-LAMP1 antibody - Lysosome Marker
BOND RX™ Validated|Lab Essentials
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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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