AB261874

人CHMP2B knockout A549 cell line

Human CHMP2B knockout A549 cell line

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CHMP2B KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%.

查看别名

ALS17, CHM2B_HUMAN, CHMP family, member 2B, CHMP2.5, Charged multivesicular body protein 2b, Chromatin-modifying protein 2b, VPS2 homolog B, VPS2B, Vacuolar protein sorting 2, yeast, homolog of, B, Vacuolar protein sorting 2-2, Vacuolar protein sorting-associated protein 2-2, Vps2-2, hVps2-2

3 Images

Lab

Western blot - Human CHMP2B knockout A549 cell line (AB261874)

Lanes 1 - 4 : Merged signal (red and green). Green - ab157208 observed at 45 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab157208 was shown to specifically react with CHMP2B in wild-type A549 cells as signal was lost in CHMP2B knockout cell line ab261874 (knockout cell lysate ab261683). Wild-type and CHMP2B knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab157208 and ab130007 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/1000 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CHMP2B antibody [EPR10807(B)] (<a href='/products/primary-antibodies/chmp2b-antibody-epr10807b-ab157208'>ab157208</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

CHMP2B knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CHMP2B knockout A549 cell line (ab261874)

Lane 3:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Predicted band size: 24 kDa

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Lab

Western blot - Human CHMP2B knockout A549 cell line (AB261874)

Lanes 1 - 4 : Merged signal (red and green). Green - ab33174 observed at 30 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab33174 was shown to recognize CHMP2B in wild-type A549 cells as signal was lost at the expected MW in CHMP2B knockout cell line ab261874 (knockout cell lysate ab261683). Additional cross-reactive bands were observed in the wild-type and knockout samples. Wild-type and CHMP2B knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% milk. ab33174 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CHMP2B antibody (<a href='/products/primary-antibodies/chmp2b-antibody-ab33174'>ab33174</a>) at 1 µg/mL

Lane 1:

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

Lane 2:

CHMP2B knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CHMP2B knockout A549 cell line (ab261874)

Lane 3:

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

Lane 4:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Predicted band size: 24 kDa

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Lab

Next Generation Sequencing - Human CHMP2B knockout A549 cell line (AB261874)

X = 1 bp insertion

关键信息

细胞类型

A549

种属

Human

组织

Lung

形式

Liquid

form

敲除验证

Next Generation Sequencing,Western blot

突变描述

Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%

疾病

Carcinoma

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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }

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产品详情

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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规格

{ "values": { "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab261874-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab261874 Human CHMP2B knockout A549 cell line", "number":"AB261874-CMP01", "productcode":"" } ] } } }

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性能和储存信息

基因名称

CHMP2B

基因编辑类型

Knockout

基因编辑方法

CRISPR technology

敲除验证

Next Generation Sequencing, Western blot

运输条件

Dry Ice

处理步骤

初始处理指南

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.

传代培养指南

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.

培养基

F-12K + 10% FBS

低温储藏试剂

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

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质量控制

STR 分析

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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细胞培养

生物安全等级

EU: 1 US: 1

贴壁/悬浮

Adherent

性别

Male

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产品实验方案

Visit the General protocols
Visit the Troubleshooting

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靶点信息

See full target information CHMP2B

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Abcam Product Promise

我们致力于为您的研究提供高质量的试剂，为您科研的每一步提供支持。若我们的产品未能达到预期性能，我们向您提供 Abcam Product Promise 保障。

详情请参阅我们的条款与条件。

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

人CHMP2B knockout A549 cell line

Human CHMP2B knockout A549 cell line

Western blot - Human CHMP2B knockout A549 cell line (AB261874)

Western blot - Human CHMP2B knockout A549 cell line (AB261874)

Next Generation Sequencing - Human CHMP2B knockout A549 cell line (AB261874)