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AB261874

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 available. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 99%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
3 Images
Western blot - Human CHMP2B knockout A549 cell line (AB261874)
  • WB

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='/en-us/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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Western blot - Human CHMP2B knockout A549 cell line (AB261874)
  • WB

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='/en-us/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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Next Generation Sequencing - Human CHMP2B knockout A549 cell line (AB261874)
  • NGS

Lab

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

X = 1 bp insertion

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

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

Disease

Carcinoma

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Complementary Products

Primary Antibodies

AB157208

Anti-CHMP2B antibody [EPR10807(B)]

RabMAb|Recombinant|KO Validated

Flow Cytometry (Intracellular) - Anti-CHMP2B antibody [EPR10807(B)] (AB157208)

  • 0
  • 0
Primary Antibodies

AB109012

Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker

KO Validated|RabMAb|Recombinant|Lab Essentials

Western blot - Anti-SQSTM1 / p62 antibody [EPR4844] - Autophagosome Marker (AB109012)

  • 5
  • 15
Primary Antibodies

AB137029

Anti-RAB7 antibody [EPR7589] - Late Endosome Marker

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Anti-RAB7 antibody [EPR7589] - Late Endosome Marker (AB137029)

  • 5
  • 24
Primary Antibodies

AB184718

Anti-MLKL antibody [EPR17514]

RabMAb|Recombinant|KO Validated|20ul selling size

Western blot - Anti-MLKL antibody [EPR17514] (AB184718)

  • 5
  • 5
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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

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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What's included?

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Properties and storage information

Gene name
CHMP2B
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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Handling procedures

Initial handling guidelines

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.

Subculture guidelines
  • 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.
Culture medium

F-12K + 10% FBS

Cryopreservation medium

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

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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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Target data

See full target information CHMP2B
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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