AB275504

Human VCAM1 knockout A549 cell lysate

Name: Human VCAM1 knockout A549 cell lysate (ab275504) | Abcam
Brand: Abcam Limited
SKU: AB275504
Price: 118000 JPY
Availability: InStock

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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.

別名を表示する

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='/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='/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='/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='/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

細胞タイプ

A549

生物種

Human

組織

Lung

ノックアウト検証方法

Immunocytochemistry,Sanger Sequencing,Western blot

ノックアウト変異

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

疾病

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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製品の詳細

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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製品内容

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出荷温度及び保存条件

遺伝子名

VCAM1

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Immunocytochemistry, Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Ambient - Can Ship with Ice

短期保存温度

-20°C

長期保存温度

-20°C

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補足情報

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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製品プロトコール

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Abcam product promise

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Human VCAM1 knockout A549 cell lysate