AB267149

Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line

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PSMB8 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 3 and 1 bp insertion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

4 Images

Lab

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Lanes 1-4 : Merged signal (red and green). Green - ab180606 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

ab180606 Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] was shown to specifically react with Proteasome 20S LMP7 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267149 (knockout cell lysate ab257130) was used. Wild-type and Proteasome 20S LMP7 knockout samples were subjected to SDS-PAGE. ab180606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and 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-Proteasome 20S LMP7 antibody [EPR14482(B)] (<a href='/en-us/products/primary-antibodies/proteasome-20s-lmp7-antibody-epr14482b-ab180606'>ab180606</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

PSMB8 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (ab267149)

Lane 3:

Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 30 kDa

Observed band size: 23 kDa

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Lab

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Lanes 1-4 : Merged signal (red and green). Green - ab82528 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

ab82528 Anti-Proteasome 20S LMP7 antibody was shown to specifically react with Proteasome 20S LMP7 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267149 (knockout cell lysate ab257130) was used. Wild-type and Proteasome 20S LMP7 knockout samples were subjected to SDS-PAGE. ab82528 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and 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-Proteasome 20S LMP7 antibody (<a href='/en-us/products/primary-antibodies/proteasome-20s-lmp7-antibody-ab82528'>ab82528</a>) at 1/500 dilution

Lane 1:

Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

PSMB8 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (ab267149)

Lane 3:

Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Lane 4:

HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Secondary

All lanes:

Predicted band size: 30 kDa

Observed band size: 23 kDa

false

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Sanger seq

Unknown

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Allele-2 : 1 bp insertion in exon 3.

Sanger seq

Unknown

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Allele-1 : 1 bp deletion in exon3

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, 1 bp deletion in exon 3 and 1 bp insertion in exon 3

Antibiotic resistance

Puromycin 1µg/mL

Disease

Carcinoma

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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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?

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab267149-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab267149 Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line", "number":"AB267149-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab255450 Human wild-type A549 cell line", "number":"AB267149-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab267149-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab267149 Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line", "number":"AB267149-CMP01", "productcode":"" } ] } } }

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Properties and storage information

Gene name

PSMB8

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Sanger Sequencing, Western blot

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.

Proteasome 20S LMP7 also known as β5i or PSMB8 is a subunit of the immunoproteasome. It weighs about 23 kDa and it is expressed predominantly in immune cells such as lymphocytes and macrophages. LMP7 is part of the proteasome 20S a cylindrical core particle involved in the degradation of ubiquitinated proteins. This process helps maintain protein homeostasis by breaking down misfolded and damaged proteins.

Biological function summary

The immunoproteasome in which LMP7 is an essential component plays an important role in the adaptive immune system. It influences the generation of antigenic peptides that are presented on MHC class I molecules. LMP7's expression is increased during immune responses particularly following gamma interferon (IFN-γ) stimulation contributing to the immune-mediated proteasome conformation. The immunoproteasome enables more efficient peptide processing enhancing antigen presentation to cytotoxic T lymphocytes.

Pathways

LMP7 is involved in the MHC class I antigen presentation pathway. This pathway allows the immune system to detect and respond to intracellular pathogens such as viruses. Additionally LMP7 interacts with TAP (Transporter associated with Antigen Processing) and other proteasome subunits to ensure effective peptide processing for loading onto MHC class I molecules. Through this involvement it closely links to immune signaling and cellular stress responses.

Research shows that LMP7 connects to autoimmune diseases and inflammatory disorders such as lupus and rheumatoid arthritis. Dysregulation of LMP7 expression or activity can lead to improper immune responses contributing to disease pathogenesis. Furthermore LMP7 impacts the degradation of specific proteins relevant to these conditions. For example it works alongside other proteasome subunits like LMP2 and MECL-1 in modulating the inflammatory response which can exacerbate autoimmune disease symptoms.

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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 PSMB8

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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 PSMB8 (Proteasome 20S LMP7) knockout A549 cell line

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267149)

Key facts

Cell type

Species or organism

Tissue

Form

Knockout validation

Mutation description

Antibiotic resistance

Disease

Complementary Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

Cell Lines & Lysates

Reactivity data

Product details

What's included?

Properties and storage information

Gene name

Gene editing type

Gene editing method

Knockout validation

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