AB261703

Human HLA-A knockout A-431 cell lysate

Name: Human HLA-A knockout A-431 cell lysate (ab261703) | Abcam
Brand: Abcam Limited
SKU: AB261703
Price: 690 USD
Availability: InStock

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HLA-A KO cell lysate available now. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control included. Knockout achieved by CRISPR/Cas9 X = 17 bp deletion Frameshift = 99.8%.

View Alternative Names

Antigen Presenting Molecule, HLA class I histocompatibility antigen, A 1 alpha chain, Leukocyte antigen class I A, MHC class I antigen HLA A heavy chain, Major histocompatibility complex, class I, A

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Supplier Data

Western blot - Human HLA-A knockout A-431 cell lysate (AB261703)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 2 : HLA A knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate 20 ug

Lane 3 : A549 (Human lung carcinoma cell line) whole cell lysate 20 ug

Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate 20 ug

Lanes 1 - 4 : Merged signal (red and green). Green - ab52922 observed at 40 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab52922 was shown to react with HLA-A in wild-type A-431 cells in Western blot Loss of signal was observed when HLA-A knockout cell line ab261894 (knockout cell lysate ab261703) was used. Wild-type A-431 and HLA-A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab52922 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed 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-HLA A antibody [EP1395Y] (<a href='/en-us/products/primary-antibodies/hla-a-antibody-ep1395y-ab52922'>ab52922</a>) at 1/10000 dilution

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

HLA A knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human HLA-A knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-hla-a-knockout-a-431-cell-line-ab261894'>ab261894</a>)

Lane 3:

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

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

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Lab

Western blot - Human HLA-A knockout A-431 cell lysate (AB261703)

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 2 : EPCAM knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lanes 1 - 3 : Merged signal (red and green). Green - ab86597 observed at 40 kDa. Red - loading control, ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55kDa.

ab86597 was shown to react with HLA-A in wild-type A-431 cells in Western blot Loss of signal was observed when HLA-A knockout cell line ab261894 (knockout cell lysate ab261703) was used. Wild-type A-431 and HLA-A knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab86597 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4°C at 5 μg/ml and a 1 in 20000 dilution respectively. Blots were developed 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:

Anti-HLA Class I A3 antibody [7g7h8] (<a href='/en-us/products/unavailable/hla-class-i-a3-antibody-7g7h8-ab86597'>ab86597</a>) at 5 µg/mL

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 2:

HLA-A knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate

Lane 2:

Western blot - Human HLA-A knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-hla-a-knockout-a-431-cell-line-ab261894'>ab261894</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

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Lab

Next Generation Sequencing - Human HLA-A knockout A-431 cell lysate (AB261703)

X = 17 bp deletion

Key facts

Cell type

A-431

Species or organism

Human

Tissue

Skin

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 17 bp deletion Frameshift = 99.8%

Disease

Epidermoid 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 and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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

HLA-A

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation 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.

HLA-A also known as Human Leukocyte Antigen A is a protein that is part of the major histocompatibility complex (MHC) class I molecules. It has an approximate molecular mass of 44 kDa. HLA-A is expressed on the surface of nearly all nucleated cells and interacts with CD8+ T lymphocytes. Known for its role in immune recognition HLA-A presents endogenous peptide antigens to the T-cell receptor on cytotoxic T cells initiating immune responses to eliminate infected or malignant cells.

Biological function summary

This target functions as a component of the MHC class I molecule complex. HLA-A binds peptides derived from proteasomal degradation of intracellular proteins and presents these peptides on the cell surface. The function plays an essential role in immune surveillance and distinction between self and non-self. This antigen presentation is important for immune responses against viral infections and tumor surveillance influencing the activation and proliferation of T lymphocytes.

Pathways

HLA-A plays a significant role in the antigen processing and presentation pathway. It is closely related to the TAP (Transporter associated with Antigen Processing) proteins which transport antigenic peptides into the endoplasmic reticulum for loading onto MHC class I molecules. Additionally the target interacts with the CD8 molecule on cytotoxic T cells. This interaction is important within the immune response pathway contributing to the recognition and destruction of infected cells.

HLA-A has been associated with autoimmune diseases and transplant rejection. It plays a significant role in conditions like ankylosing spondylitis where specific HLA-A subtypes contribute to disease susceptibility. Additionally mismatches in HLA-A typing can lead to transplant rejection. In such scenarios anti-HLA antibodies can develop causing damage to transplant tissues. The complex interaction of HLA-A with related proteins such as other MHC class I molecules influences immune response outcomes in these 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

Female

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Product protocols

Visit the General protocols
Visit the Troubleshooting

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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 HLA-A knockout A-431 cell lysate

Western blot - Human HLA-A knockout A-431 cell lysate (AB261703)

Western blot - Human HLA-A knockout A-431 cell lysate (AB261703)

Next Generation Sequencing - Human HLA-A knockout A-431 cell lysate (AB261703)

Key facts

Cell type

Species or organism

Tissue

Knockout validation

Mutation description

Disease

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

Supplementary information

Biological function summary

Pathways

Quality control

STR analysis

Cell culture

Biosafety level

Adherent/suspension

Gender

Product protocols

Complementary Products

Cell Lines & Lysates

Primary Antibodies

Primary Antibodies

Primary Antibodies

Product promise