AB273748

Human BSG (CD147) knockout A549 cell line

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BSG KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 78 bp insertion in exon 5 introducing premature STOP codon. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

11 Images

Flow Cytometry (Intracellular) - Human BSG (CD147) knockout A549 cell line (AB273748)

Flow Cyt (Intra)

Lab

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 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 (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) (ab150117) was used at 1/2000 for 30 min at 4°C.

Isotype control antibody was mouse IgG1κ (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.

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

Flow Cyt

Lab

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 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 (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) (ab150117) was used at 1/2000 for 30 min at 4°C.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Flow Cyt

Lab

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 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 (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) (ab150117) was used at 1/2000 for 30 min at 4°C.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

Flow Cyt

Lab

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 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 (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) (ab150117) was used at 1/2000 for 30 min at 4°C.

Isotype control antibody was mouse IgG2bκ (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.

Lab

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Lanes 1 - 4 : Merged signal (red and green). Green - ab666 observed at 55-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

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 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 ab666 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (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] (<a href='/en-us/products/primary-antibodies/cd147-antibody-mem-m6-1-ab666'>ab666</a>) 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

Predicted band size: 42 kDa

Observed band size: 55-70 kDa

false

Lab

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Lanes 1 - 4 : Merged signal (red and green). Green - ab119020 observed at 55-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

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 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 ab119020 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD147 antibody [OTI9B10] (<a href='/en-us/products/primary-antibodies/cd147-antibody-oti9b10-ab119020'>ab119020</a>) 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

Predicted band size: 42 kDa

Observed band size: 55-70 kDa

false

Lab

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Lanes 1 - 3 : Merged signal (red and green). Green - ab108308 observed at 42-70 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

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 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 ab108308 and 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD147 antibody [EPR4053] (<a href='/en-us/products/primary-antibodies/cd147-antibody-epr4053-ab108308'>ab108308</a>) 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

Predicted band size: 42 kDa

Observed band size: 42-70 kDa

false

Lab

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Lanes 1 - 4 : Merged signal (red and green). Green - ab230921 observed at 42-70 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

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 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 ab230921 and 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 (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD147 antibody [10E10] (<a href='/en-us/products/primary-antibodies/cd147-antibody-10e10-ab230921'>ab230921</a>) 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

Predicted band size: 42 kDa

Observed band size: 42-70 kDa

false

Flow Cyt

Lab

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 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 (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) (ab150117) was used at 1/2000 for 30 min at 4°C.

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)

sELISA

Lab

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 (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)

Sanger seq

Lab

Sanger Sequencing - Human BSG (CD147) knockout A549 cell line (AB273748)

Allele-1 : 78 bp insertion in exon 5 introducing premature STOP codon.

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "Flow Cyt": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

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Product details

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.

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What's included?

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Properties and storage information

Gene name

BSG

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

Zygosity

Homozygous

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

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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.

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Supplementary information

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

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.

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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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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Target data

See full target information BSG

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Human BSG (CD147) knockout A549 cell line

Flow Cytometry (Intracellular) - Human BSG (CD147) knockout A549 cell line (AB273748)

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

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

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

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

Western blot - Human BSG (CD147) knockout A549 cell line (AB273748)

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

Sandwich ELISA - Human BSG (CD147) knockout A549 cell line (AB273748)

Sanger Sequencing - Human BSG (CD147) knockout A549 cell line (AB273748)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Disease

Complementary Products

Cell Lines & Lysates

Cell Lines & Lysates

Cell Lines & Lysates

Cell Lines & Lysates

Reactivity data

Product details

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

Subculture guidelines

Culture medium

Cryopreservation medium

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Target data

Product promise