Human CCL4 knockout THP-1 cell line
- Advanced Validation
- What is this?
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CCL4L1 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: 7 bp deletion in exon 1.
View Alternative Names
ACT-2, AT744.1, AT744.2, C C motif chemokine 4, C C motif chemokine 4 like, C C motif chemokine ligand 4 like 1, C C motif chemokine ligand 4 like 2, CC chemokine ligand 4, CC chemokine ligand 4L1, CC chemokine ligand 4L1d2, CC chemokine ligand 4L2, CCL4L, CCL4_HUMAN, Chemokine (C C motif) ligand 4, Chemokine (C C motif) ligand 4 like 1, Chemokine (C C motif) ligand 4 like 1, telomeric, Chemokine (C C motif) ligand 4 like 2, Chemokine CC Motif Ligand 4, G 26, G-26 T-lymphocyte-secreted protein, HC21, Immune activation 2, LAG-1, Lymphocyte activation gene 1, Lymphocyte activation gene 1 protein, MGC104418, MGC126025, MGC126026, MIP-1-beta, MIP-1-beta(1-69), MIP-1-beta(3-69), MIP1B, MIP1B1, Macrophage inflammatory protein 1-beta, Macrophage inflammatory protein 1b2, Monocyte adherence induced protein 5 alpha, PAT 744, Protein H400, SCYA2, SCYA4, SCYA4L, SCYA4L1, SCYA4L2, SCYQ4L2, SIS-gamma, Secreted protein G 26, Small inducible cytokine A4 (homologous to mouse Mip 1b), Small-inducible cytokine A4, T-cell activation protein 2, ccl4l 1, small inducible cytokine A4-like
- WB
Lab
Western blot - Human CCL4 knockout THP-1 cell line (AB273719)
False colour image of Western blot : Anti-CCL4/MIP-1 beta antibody [EP521Y] staining at 1/1000 dilution shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot ab45690 was shown to bind specifically to CCL4/MIP-1 beta. A band was observed at 12 kDa in wild-type THP-1 cell lysates with no signal observed at this size in CCL4 knockout cell line ab273719 (knockout cell lysate ab275512). To generate this image wild-type and CCL4 knockout THP-1 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 °. 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-CCL4/MIP-1 beta antibody [EP521Y] (<a href='/en-us/products/primary-antibodies/ccl4-mip-1-beta-antibody-ep521y-ab45690'>ab45690</a>) at 1/1000 dilution
Lane 1:
Wild-type THP-1 Vehicle control + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Lane 2:
Wild-type THP-1 Treated PMA (100 ng/mL, 56 h) + LPS (1 u/mL, 24 h) + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Lane 2:
Western blot - Human CCL4 knockout THP-1 cell line (ab273719)
Lane 3:
CCL4 knockout THP-1 Vehicle control + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Lane 4:
CCL4 knockout THP-1 Treated PMA (100 ng/mL, 56 h) + LPS (1 u/mL, 24 h) + Brefeldin A (5 u/mL, 6 h) cell lysate at 20 µg
Predicted band size: 10 kDa
Observed band size: 12 kDa
false
- NGS
Lab
Next Generation Sequencing - Human CCL4 knockout THP-1 cell line (AB273719)
Allele-1 : 7 bp deletion in exon 1
Reactivity data
Product details
Recommended control: Human wild-type THP-1 cell line (ab281894). 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.
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Upon thawing make banks as soon as possible and use each bank within 10-20 passages
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As cell line growth can vary please attempt culture for 2 weeks from revival of initial bank before contacting the technical team.
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.
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. Based on cell count, seed cells in an appropriate cell culture flask at a density of 4x105 cells/mL. Seeding density is given as a guide only and should be scaled to align with individual lab schedules.
4. Incubate the culture at 37°C incubator with 5% CO2. Cultures should be monitored daily.
5. THP-1 cells recover slowly from cryopreservation and therefore may not be ready for subculture for a number of days. Cells should be left as much as possible over this time and only subcultured when the cell density reaches 8x105 cells/mL.Small amounts of fresh media can be added until cell number/viability improves.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- Cells should be seeded at 2x105 - 3x105 cells/mL and subcultured when they have reached 8x105 cells/mL.
- It is not recommended to allow the cell density to exceed 1x106 cells/mL.
Culture medium
RPMI + 10% FBS + 0.05 mM beta-mercaptoethanol
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
CCL4 is critical in immune system modulation. It coordinates leukocyte activation and trafficking contributing to inflammation and immune surveillance. It does not function in isolation but rather interacts with other chemokines and receptors. CCL4 alongside its sibling chemokines forms a dynamic network that ensures a balanced immune response. It connects to receptors such as CCR5 playing substantial roles in its activities.
Pathways
CCL4 participates in essential signaling cascades that mediate immune responses. It is notably part of the chemokine signaling pathway which is instrumental in directing cell movement. CCL4 also engages with proteins like CCR5 in the inflammatory and immune response pathways. This interaction enhances communication between cells required for effective pathogen defense.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Suspension
Gender
Male
Target data
Complementary Products
Cell Lines & Lysates
AB276116
Human CASP1 (Caspase-1) knockout THP-1 cell line
Advanced Validation
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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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