Human CCL2 (MCP1) knockout A549 cell line
- Advanced Validation
- What is this?
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CCL2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift: 97.75%.
View Alternative Names
C-C motif chemokine 2, CCL2_HUMAN, Chemokine (C C motif) ligand 2, GDCF-2, HC11, HSMCR30, JE, MCAF, MCP-1, MGC9434, Monocyte chemoattractant protein 1, Monocyte chemotactic and activating factor, Monocyte chemotactic protein-1, Monocyte secretory protein JE, SCYA2, SMC-CF, Small inducible cytokine A2 (monocyte chemotactic protein 1, homologous to mouse Sig je), Small inducible cytokine subfamily A (Cys Cys), member 2, Small-inducible cytokine A2
- WB
Lab
Western blot - Human CCL2 (MCP1) knockout A549 cell line (AB270478)
Anti-CCL2 antibody [EPR21025] (ab214819) staining at 1/400 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab214819 was shown to bind specifically to CCL2. A band was observed at 13 kDa in wild-type A549 cell lysates with no signal observed at this size in CCL2 knockout cell line ab270478 (knockout cell lysate ab270501). To generate this image, wild-type and CCL2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 at 1/20000 dilution and Goat anti-Mouse IgG H&L 680RD at 1/80000 dilution
All lanes:
Western blot - Anti-MCP1 antibody [EPR21025] (<a href='/en-us/products/primary-antibodies/mcp1-antibody-epr21025-ab214819'>ab214819</a>)
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
Wild-type A549 Treated BFA (1 ug/mL, 3 h) cell lysate, at 20 µg
Lane 3:
CCL2 knockout A549 Treated BFA (1 ug/mL, 3 h) cell lysate at 20 µg
Observed band size: 13 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human CCL2 (MCP1) knockout A549 cell line (AB270478)
Knockout achieved by CRISPR/Cas9; X = 1 bp insertion; Frameshift : 97.75%
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.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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
Shipped at conditions
Appropriate short-term storage duration
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
The role of MCP-1 is significant in guiding immune cells to areas requiring immune response. It does not form part of a complex but operates as a singular entity to affect immune signaling. MCP-1 influences the behavior of monocytes and macrophages by inducing chemotaxis and activating their effector functions. Through this mechanism it aids in the inflammatory response and tissue repair.
Pathways
MCP-1 participates in the inflammatory pathways by cooperating with key proteins like CCR2. It facilitates the JAK-STAT signaling pathway which is vital for transmitting extracellular information into cellular responses. The interaction of MCP-1 with CCR2 influences the activation of downstream proteins like STAT3 enabling various immune responses. MCP-1 thereby plays a central role in modulating inflammation-related pathways.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Complementary Products
Primary Antibodies
AB214819
BOND RX™ Validated|RabMAb|Recombinant|KO Validated
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MCP1 antibody [EPR21025] (AB214819)](https://content.abcam.com/products/images/mcp1-antibody-epr21025-ab214819--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img428945.jpg)
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Product promise
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