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AB273760

Human CCL3 (Macrophage Inflammatory Protein 1) knockout THP-1 cell line

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CCL3 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 61 bp deletion in exon 1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
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Sanger Sequencing - Human CCL3 (Macrophage Inflammatory Protein 1) knockout THP-1 cell line (AB273760)
  • Sanger seq

Lab

Sanger Sequencing - Human CCL3 (Macrophage Inflammatory Protein 1) knockout THP-1 cell line (AB273760)

Allele-1 : 61 bp deletion in exon 1

Key facts

Cell type

THP-1

Species or organism

Human

Tissue

Blood

Form

Liquid

form

Knockout validation

Sanger Sequencing

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 61 bp deletion in exon 1

Disease

Acute Monocytic Leukemia

Product details

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?

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Properties and storage information

Gene name
CCL3
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing
Zygosity
Homozygous
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C

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.

MIP-1 alpha also known as CCL3 is a chemokine that plays a critical role in the immune system. With a molecular weight of around 8 kDa it is primarily expressed in activated T-cells macrophages and fibroblasts. MIP-1 alpha acts as a signaling protein that binds to receptors on target cells directing the movement of immune cells. Researchers commonly refer to it using various alternative names such as MIP-1a and CCL3-alpha rat and it is frequently measured using assays like MIP-1 alpha ELISA for experimental analyses.
Biological function summary

Proteins including MIP-1 alpha participate in the recruitment and activation of diverse immune cells aiding the inflammatory response. Through interaction with receptors like CCR1 and CCR5 MIP-1 alpha mobilizes cells like natural killer cells and monocytes. This chemokine does not generally form part of a larger protein complex but interacts directly with membrane receptors to exert its function in immune cell chemotaxis.

Pathways

MIP-1 alpha operates in the context of both the inflammatory response and the chemokine signaling pathways. Its involvement in these pathways helps mediate the trafficking of leukocytes. Notably MIP-1 alpha interacts with proteins such as CCR1 and CCR5 to transmit signals that instigate immune responses. These proteins are integral to its role in modulating immunity providing targets for research into controlling inflammatory processes.

MIP-1 alpha closely relates to conditions such as rheumatoid arthritis and HIV infection. In rheumatoid arthritis its overexpression can lead to excessive inflammation and joint destruction. Meanwhile in HIV infection MIP-1 alpha interacts with CCR5 a co-receptor necessary for the virus entry into cells. This dual relationship elucidates its potential as a therapeutic target in managing both inflammatory diseases and viral entry mechanisms.

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Suspension

Gender

Male

Product protocols

Product promise

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