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AB267148

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 provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 3.

View Alternative Names

ALDD, D6S216, D6S216E, LMP 7, Large multifunctional peptidase 7, Large multifunctional protease 7, Low molecular mass protein 7, Low molecular weight protein 7, MGC1491, Macropain subunit C13, Multicatalytic endopeptidase complex subunit C13, NKJO, OTTHUMP00000062981, PSB8_HUMAN, PSMB 8, PSMB5i, Protease component C13, Proteasome (prosome macropain) subunit beta type 8, Proteasome (prosome, macropain) subunit, beta type, 8 (large multifunctional peptidase 7), Proteasome beta 8 subunit, Proteasome catalytic subunit 3i, Proteasome component C13, Proteasome related gene 7, Proteasome subunit Y2, Proteasome subunit beta 8, Proteasome subunit beta type, Proteasome subunit beta type-8, Proteasome subunit beta-5i, RING 10, Really interesting new gene 10 protein, Y2

4 Images
Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)
  • WB

Unknown

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)

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 ab267148 (knockout cell lysate ab257129) 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°C 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 cell lysate at 20 µg

Lane 2:

PSMB8 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (ab267148)

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HEK-293 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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Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)
  • WB

Lab

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)

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 ab267148 (knockout cell lysate ab257129) 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 overnight at 4°C 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 cell lysate at 20 µg

Lane 2:

PSMB8 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (ab267148)

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HEK-293 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

false

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)
  • Sanger seq

Unknown

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)

Homozygous : 1 bp insertion in exon3

Cell Culture - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)
  • Cell Culture

Unknown

Cell Culture - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell line (AB267148)

Representative images of PSMB8 knockout A549 cells, low and high confluency examples (top left and right respectively) and wild-type A549 cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using an EVOS M5000 microscope.

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: 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

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.

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What's included?

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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
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.

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

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Target data

See full target information PSMB8
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Complementary Products

Primary Antibodies

AB180606

Anti-Proteasome 20S LMP7 antibody [EPR14482(B)]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (AB180606)

  • 5
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Primary Antibodies

AB166628

Anti-Proteasome subunit beta type 2/PSMB2 antibody [EPR8826(2)(B)]

RabMAb|Recombinant

Western blot - Anti-Proteasome subunit beta type 2/PSMB2 antibody [EPR8826(2)(B)] (AB166628)

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  • 5
Primary Antibodies

AB22673

Anti-Proteasome 20S alpha + beta antibody

Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S alpha + beta antibody (AB22673)

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  • 7
Cell Lines & Lysates

AB266765

Human ADRM1 (ARM-1) knockout HEK-293T cell line

Advanced Validation

Western blot - Human ADRM1 (ARM-1) knockout HEK-293T cell line (AB266765)

  • 0
  • 0

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