AB275504

Human VCAM1 knockout A549 cell lysate

Name: Human VCAM1 knockout A549 cell lysate (ab275504) | Abcam
Brand: Abcam Limited
SKU: AB275504

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VCAM1 KO cell lysate available now. KO validated by Immunocytochemistry, Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 2.

View Alternative Names

CD106, CD106 Antigen, INCAM-100, MGC99561, VCAM1_HUMAN, Vascular Cell Adhesion Molecule 1, Vascular cell adhesion protein 1

3 Images

Lab

Western blot - Human VCAM1 knockout A549 cell lysate (AB275504)

Lane 1 : Wild-type A549 cell lysate 30 ug
Lane 2 : Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate 30 ug
Lane 3 : VCAM1 knockout A549 cell lysate 30 ug
Lane 4 : VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate 30 ug
Lane 5 : HUVEC cell lysate 30 ug
Lane 6 : HUVEC TNF-a treated (16 ng/mL, 16h) cell lysate 30 ug
Lanes 1 - 6 : Merged signal (red and green). Green - ab134047 observed at 105 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab134047 was shown to react with VCAM1 in wild-type A549 cells in Western blot with loss of signal observed in VCAM1 knockout cell line ab273758 (knockout cell lysate ab275504). Wild-type A549 and VCAM1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab134047 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 ^°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-VCAM1 antibody [EPR5047] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr5047-ab134047'>ab134047</a>) at 1/2000 dilution

Lane 1:

Wild-type A549 cell lysate at 30 µg

Lane 2:

Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg

Lane 2:

Western blot - Human VCAM1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-vcam1-knockout-a549-cell-line-ab273758'>ab273758</a>)

Lane 3:

VCAM1 knockout A549 cell lysate at 30 µg

Lane 4:

VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg

Lane 5:

HUVEC cell lysate at 30 µg

Lane 6:

HUVEC TNF-a treated (16 ng/mL, 16h) cell lysate at 30 µg

Predicted band size: 81 kDa

Observed band size: 105 kDa

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Lab

Western blot - Human VCAM1 knockout A549 cell lysate (AB275504)

Lanes 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 2 : GPX4 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate, 20 ug
Lanes 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate, 20 ug
Lanes 3 : Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate, 20 ug

ab174279 was shown to react with VCAM1 in treated wild-type A549 cells in western blot. Loss of signal was observed when treated VCAM1 knockout cell line ab273758 (knockout cell lysate ab275504) was used. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab174279 overnight at 4 °C at a 1 in 1000 dilution. Blots were incubated with HRP conjugated Goat anti-Rabbit (H+L) secondary antibody at 1 in 5000 for 1 hour at room temperature before development with Optiblot ECL reagent (ab133456) and imaging.

All lanes:

Western blot - Anti-VCAM1 antibody [EPR5038(2)] (<a href='/en-us/products/primary-antibodies/vcam1-antibody-epr50382-ab174279'>ab174279</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 30 µg

Lane 2:

Wild-type A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg

Lane 3:

VCAM1 knockout A549 cell lysate at 30 µg

Lanes 3 - 4:

Western blot - Human VCAM1 knockout A549 cell line (<a href='/en-us/products/cell-lines/human-vcam1-knockout-a549-cell-line-ab273758'>ab273758</a>)

Lane 4:

VCAM1 knockout A549 TNF-a treated (10 ng/mL, 16h) cell lysate at 30 µg

Predicted band size: 81 kDa

Observed band size: 105 kDa

false

Exposure time: 20s

Sanger Sequencing - Human VCAM1 knockout A549 cell lysate (AB275504)

Sanger seq

Lab

Sanger Sequencing - Human VCAM1 knockout A549 cell lysate (AB275504)

Allele-1 : 4 bp deletion in exon 2

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Knockout validation

Immunocytochemistry,Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, Homozygous: 4 bp deletion in exon 2

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.

Treatments:
Human VCAM1 knockout A549 cell lysate - Untreated
Human wild-type A549 cell lysate - Untreated
Human VCAM1 knockout A549 cell lysate - TNF-a (10 ng/ml, 16h)
Human wild-type A549 cell lysate - TNF-a (10 ng/ml, 16h)

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

VCAM1

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Immunocytochemistry, Sanger Sequencing, Western blot

Zygosity

Homozygous

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.

VCAM1 also known as vascular cell adhesion molecule 1 plays an important role in mediating adhesion and signal transduction. VCAM1 has an approximate molecular mass of 110 kDa. It is expressed on the surface of endothelial cells and can be upregulated by cytokines released during inflammation. The protein serves as a ligand for the integrin VLA-4 also known as α4β1 and contributes to the adhesion of leukocytes to endothelial cells.

Biological function summary

Adhesion molecules like VCAM1 participate in immune response by recruiting leukocytes to sites of inflammation. VCAM1 helps guide immune cells to directed locations where they perform defense activities. Although VCAM1 is not part of any known large protein complex its interaction with integrins facilitates the migration of immune cells across the endothelium and into tissue.

Pathways

VCAM1 contributes significantly to the leukocyte extravasation process in inflammatory pathways. It plays a role in both the cytokine-cytokine receptor interaction pathway and the NF-kB signaling pathway which are essential for immune response and cell signaling. Through these pathways it interacts with proteins like ICAM1 and various chemokines helping to coordinate the immune cell movement through vascular tissue barriers.

Abnormal regulation of VCAM1 links to various cardiovascular diseases such as atherosclerosis and inflammatory diseases like rheumatoid arthritis. The protein levels may increase in these conditions promoting excessive immune cell recruitment and tissue damage. VCAM1 also interfaces with other proteins like ICAM1 in these pathological states which together contribute to the progression and severity of 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

Male

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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 VCAM1 knockout A549 cell lysate

Western blot - Human VCAM1 knockout A549 cell lysate (AB275504)

Western blot - Human VCAM1 knockout A549 cell lysate (AB275504)

Sanger Sequencing - Human VCAM1 knockout A549 cell lysate (AB275504)

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

Zygosity

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

Primary Antibodies

Primary Antibodies

Cell Lines & Lysates

Assay Kits

Product promise