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AB257129

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

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PSMB8 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

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

3 Images
Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell lysate (AB257129)
  • WB

Unknown

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell lysate (AB257129)

Lane 1 : Wild-type A549 cell lysate (20 ug)
Lane 2 : PSMB8 knockout A549 cell lysate (20 ug)
Lane 3 : Raji cell lysate (20 ug)
Lane 4 : HEK-293 cell lysate (20 ug)

ab82528 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 4oC 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 (<a href='/en-us/products/cell-lines/human-psmb8-proteasome-20s-lmp7-knockout-a549-cell-line-ab267148'>ab267148</a>)

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 lysate (AB257129)
  • WB

Lab

Western blot - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell lysate (AB257129)

Lane 1 : Wild-type A549 cell lysate (20 ug)
Lane 2 : PSMB8 knockout A549 cell lysate (20 ug)
Lane 3 : Raji cell lysate (20 ug)
Lane 4 : HEK-293 cell lysate (20 ug)

ab180606 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 4oC 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 (<a href='/en-us/products/cell-lines/human-psmb8-proteasome-20s-lmp7-knockout-a549-cell-line-ab267148'>ab267148</a>)

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 lysate (AB257129)
  • Sanger seq

Unknown

Sanger Sequencing - Human PSMB8 (Proteasome 20S LMP7) knockout A549 cell lysate (AB257129)

Homozygous : 1 bp insertion in exon3

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp insertion in exon3.

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

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.

User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.

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": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257129-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human PSMB8 knockout A549 cell lysate", "number":"AB257129-CMP02", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type A549 cell lysate", "number":"AB257129-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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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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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Product promise

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