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AB255432

Human SMARCA4 (BRG1) knockout HEK-293T cell line

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SMARCA4 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
5 Images
Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)
  • WB

Lab

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)

Lanes 1- 2 : Merged signal (red and green). Green - ab110641 observed at 185 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab110641 was shown to react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate ab263853) was used. Wild-type HEK-293T and SMARCA4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab110641 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-BRG1 antibody [EPNCIR111A] (<a href='/en-us/products/primary-antibodies/brg1-antibody-epncir111a-ab110641'>ab110641</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SMARCA4 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)

Predicted band size: 185 kDa

Observed band size: 185 kDa

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Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)
  • WB

Lab

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)

Lanes 1- 2 : Merged signal (red and green). Green - ab108318 observed at 185 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

ab108318 was shown to react with SMARCA4 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255432 (knockout cell lysate ab263853) was used. Wild-type HEK-293T and SMARCA4 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab108318 and Anti-Vinculin antibody [VIN-54] 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-BRG1 antibody [EPR3912] (<a href='/en-us/products/primary-antibodies/brg1-antibody-epr3912-ab108318'>ab108318</a>) at 1/10000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SMARCA4 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)

Predicted band size: 185 kDa

Observed band size: 185 kDa

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Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)
  • WB

Lab

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)

False colour image of Western blot : Anti-BRG1 antibody [EPNCIR111A] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab110641 was shown to bind specifically to BRG1. A band was observed at 185 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in SMARCA4 knockout cell line ab255432 (knockout cell lysate ab263853). To generate this image, wild-type and SMARCA4 knockout HEK-293T cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

All lanes:

Western blot - Anti-BRG1 antibody [EPNCIR111A] (<a href='/en-us/products/primary-antibodies/brg1-antibody-epncir111a-ab110641'>ab110641</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

SMARCA4 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell line (ab255432)

Predicted band size: 185 kDa

Observed band size: 185 kDa

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Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)

Allele-2 : 1 bp deletion in exon 4.

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell line (AB255432)

Allele-1 : 7 bp deletion in exon 4

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4

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Complementary Products

Primary Antibodies

AB110641

Anti-BRG1 antibody [EPNCIR111A]

BOND RX™ Validated|Recombinant|KO Validated|Advanced Validation|RabMAb

ChIC/CUT&RUN sequencing - Anti-BRG1 antibody [EPNCIR111A] (AB110641)

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Primary Antibodies

AB222519

Anti-SNF5/SMARCB1 antibody [EPR20189]

RabMAb|Recombinant|Trial Size

Flow Cytometry (Intracellular) - Anti-SNF5/SMARCB1 antibody [EPR20189] (AB222519)

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  • 0
Primary Antibodies

AB240648

Anti-SMARCA2 / BRM antibody [EPR23103-44]

BOND RX™ Validated|RabMAb|Recombinant|KO Validated|20ul selling size

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMARCA2 / BRM antibody [EPR23103-44] (AB240648)

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Cell Lines & Lysates

AB263853

Human SMARCA4 (BRG1) knockout HEK-293T cell lysate

Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (AB263853)

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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 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
SMARCA4
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger 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.
Culture medium

DMEM (High Glucose) + 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: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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Target data

See full target information SMARCA4
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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