Human BSG (CD147) knockout A549 cell line (ab273748)

BSG 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: 78 bp insertion in exon 5 introducing premature STOP codon.

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Images

Flow Cytometry - Human BSG (CD147) knockout A549 cell line (AB273748), expandable thumbnail

Key facts

Cell type: A549
Species or organism: Human
Tissue: Lung
Form: Liquid
Knockout validation: Sanger Sequencing, Western blot
Mutation description: Knockout achieved by using CRISPR/Cas9, Homozygous: 78 bp insertion in exon 5 introducing premature STOP codon

Alternative names

5A11 antigen, 5F7, BASI_HUMAN, Basigin, Basigin (Ok blood group), Blood brain barrier HT7 antigen, Bsg, CD 147, CD147 antigen, Collagenase stimulatory factor, EMMPRIN, Extracellular matrix metalloproteinase inducer, Leukocyte activation antigen M6, M 6, M6 leukocyte activation antigen, Neurothelin, OK, OK blood group, OK blood group antigen, TCSF, Tumor cell-derived collagenase stimulatory factor

Key facts

Cell type: A549
Species or organism: Human
Tissue: Lung
Form: Liquid
Knockout validation: Sanger Sequencing, Western blot
Mutation description: Knockout achieved by using CRISPR/Cas9, Homozygous: 78 bp insertion in exon 5 introducing premature STOP codon
Disease: Carcinoma
Concentration

Properties

Gene name: BSG
Gene editing type: Knockout
Gene editing method: CRISPR technology
Knockout validation: Sanger Sequencing, Western blot
Zygosity: Homozygous

Quality control

STR analysis: CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level: EU: 1 US: 1
Adherent/suspension: Adherent
Gender: Male

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.

Storage

Shipped at conditions: Dry Ice
Appropriate short-term storage conditions: -196°C
Appropriate long-term storage conditions: -196°C

Notes

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD147 also referred to as EMMPRIN or basigin is a transmembrane glycoprotein with a molecular weight of roughly 50–60 kDa. This protein is found on the surface of many cell types including leukocytes platelets and endothelial cells and it plays a critical role in cell function related to immune responses and cellular interactions. CD147 interacts with several cellular components serving mechanical functions such as facilitating cell-to-cell communication and contributing to the stability of cell structures. The protein is ubiquitously expressed but exhibits higher levels in tissues like the brain skin and liver.

Biological function summary

CD147 engages in multiple roles beyond its mechanical functions. It functions as a part of protein complexes and is involved in processes such as regulation of matrix metalloproteinases (MMPs) which are pivotal in tissue remodeling and repair. It can modulate inflammatory responses and is essential for cellular signaling pathways that affect cellular metabolism and growth. The involvement of CD147 in immune responses indicates its importance in various physiological processes.

Pathways

CD147 plays a significant role in the MAPK and NF-kB signaling pathways which are fundamental for controlling inflammatory responses and cell survival. It interacts with proteins such as integrins and cyclophilins which are important for mediating these pathways. These interactions highlight CD147's impact on cellular dynamics and its potential role in modulating the cellular environment in response to external stimuli.

Associated diseases and disorders

CD147 has been implicated in cancer promotion and progression as well as inflammatory diseases like rheumatoid arthritis. Its overexpression is often observed in tumors where it interacts with MMPs to facilitate tumor invasion and metastasis. In inflammatory conditions CD147 can influence the behavior of proteins like cytokines exacerbating symptoms and contributing to disease severity. These associations make CD147 a potential target for therapeutic intervention in cancer and inflammatory diseases.

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11 product images

Flow Cytometry - Human BSG (CD147) knockout A549 cell line (ab273748)
Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with Anti-CD147 antibody [MEM-M6/2] ab21903 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-CD147 antibody [MEM-M6/2] ab21903) (1x10⁶ in 100μl at 5 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG2bκ (Mouse IgG2b, kappa monoclonal [7E10G10] - Isotype Control ab170192) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Flow Cytometry - Human BSG (CD147) knockout A549 cell line (ab273748)
Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with Anti-CD147 antibody [MEM-M6/1] ab666 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-CD147 antibody [MEM-M6/1] ab666) (1x10⁶ in 100μl at 10 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG1κ (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD147 antibody [OTI9B10] ab119020 observed at 55-70 kDa. Red - loading control Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Anti-CD147 antibody [OTI9B10] ab119020 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate Human BSG (CD147) knockout A549 cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-CD147 antibody [OTI9B10] ab119020 and Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4° at a 1 in 2000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CD147 antibody [OTI9B10] (Anti-CD147 antibody [OTI9B10] ab119020) at 1/2000 dilution
Lane 1: Wild-type A549 cell lysate at 30 µg
Lane 2: BSG knockout A549 cell lysate at 30 µg
Lane 2: Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lane 3: Raji cell lysate at 30 µg
Lane 4: Jurkat cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 55-70 kDa
Flow Cytometry - Human BSG (CD147) knockout A549 cell line (ab273748)
This data was developed using the same antibody clone in a different buffer formulation (Anti-CD147 antibody [8D6] ab194401)
Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with Anti-CD147 antibody [8D6] ab194401 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-CD147 antibody [8D6] ab194401) (1x10⁶ in 100μl at 10 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG1κ (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lanes 1 - 3: Merged signal (red and green). Green - Anti-CD147 antibody [EPR4053] ab108308 observed at 42-70 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
Anti-CD147 antibody [EPR4053] ab108308 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate Human BSG (CD147) knockout A549 cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-CD147 antibody [EPR4053] ab108308 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4° at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (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-CD147 antibody [EPR4053] (Anti-CD147 antibody [EPR4053] ab108308) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 30 µg
Lane 2: BSG knockout A549 cell lysate at 30 µg
Lane 2: Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lane 3: Raji cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42-70 kDa
Flow Cytometry - Human BSG (CD147) knockout A549 cell line (ab273748)
Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with Anti-CD147 antibody [VJ1/9] ab91147 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-CD147 antibody [VJ1/9] ab91147) (1x10⁶ in 100μl at 10 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG1κ (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD147 antibody [MEM-M6/1] ab666 observed at 55-70 kDa. Red - loading control Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Anti-CD147 antibody [MEM-M6/1] ab666 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate Human BSG (CD147) knockout A549 cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-CD147 antibody [MEM-M6/1] ab666 and Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4° at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CD147 antibody [MEM-M6/1] (Anti-CD147 antibody [MEM-M6/1] ab666) at 1 µg/mL
Lane 1: Wild-type A549 cell lysate at 30 µg
Lane 2: BSG knockout A549 cell lysate at 30 µg
Lane 2: Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lane 3: Raji cell lysate at 30 µg
Lane 4: Jurkat cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 55-70 kDa
Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-CD147 antibody [10E10] ab230921 observed at 42-70 kDa. Red - loading control Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.
Anti-CD147 antibody [10E10] ab230921 was shown to react with CD147 in wild-type A549 cells in western blot with loss of signal observed in BSG knockout cell line ab273748 (knockout cell lysate Human BSG (CD147) knockout A549 cell lysate ab275500). Wild-type and BSG knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with Anti-CD147 antibody [10E10] ab230921 and Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4° at a 1 in 500 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-CD147 antibody [10E10] (Anti-CD147 antibody [10E10] ab230921) at 1/500 dilution
Lane 1: Wild-type A549 cell lysate at 30 µg
Lane 2: BSG knockout A549 cell lysate at 30 µg
Lane 2: Western blot - Human BSG (CD147) knockout A549 cell line (ab273748)
Lane 3: Raji cell lysate at 30 µg
Lane 4: Jurkat cell lysate at 30 µg
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42-70 kDa
Flow Cytometry (Intracellular) - Human BSG (CD147) knockout A549 cell line (ab273748)
Flow cytometry overlay histogram showing wild-type A549 (green line) and BSG knockout A549 cells (ab273748) stained with Anti-CD147 antibody [10E10] ab230921 (red line). The cells were incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (Anti-CD147 antibody [10E10] ab230921) (1x10⁶ in 100μl at 5 μg/ml) for 30 min at 4°C.
The secondary antibody Goat anti-mouse IgG H&L (Alexa Fluor^® 488, pre-adsorbed) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) was used at 1/2000 for 30 min at 4°C.
Isotype control antibody was mouse IgG1κ (Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control ab170190) used at the same concentration and conditions as the primary antibody (wild-type A549 - black line; BSG knockout A549 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
Sandwich ELISA - Human BSG (CD147) knockout A549 cell line (ab273748)
Human CD147 concentration was interpolated from the EMMPRIN (CD147) standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human EMMPRIN ELISA kit (Human EMMPRIN ELISA Kit ab219631). Wild-type and CD147 knockout A549 cells (ab273748) were assessed in duplicate (n=2); Jurkat and Raji cells were used as positive and negative controls respectively (n=1). Where samples were run in duplicate, data are represented as the mean and error bars represent standard deviation.
Sanger Sequencing - Human BSG (CD147) knockout A549 cell line (ab273748)
Allele-1: 78 bp insertion in exon 5 introducing premature STOP codon.