JavaScript is disabled in your browser. Please enable JavaScript to view this website.
AB263853

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

Be the first to review this product! Submit a review

|

(0 Publication)

SMARCA4 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 4 and 7 bp deletion in exon 4.
5 Images
Western blot - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (AB263853)
  • WB

Lab

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

Lane 1 : Wild-type HEK-293T cell lysate (20 ug)
Lane 2 : SMARCA4 knockout HEK-293T cell lysate (20 ug)

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 (<a href='/en-us/products/cell-lines/human-smarca4-brg1-knockout-hek-293t-cell-line-ab255432'>ab255432</a>)

Predicted band size: 185 kDa

Observed band size: 185 kDa

false

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

Lab

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

Lane 1 : Wild-type HEK-293T cell lysate (20µg)

Lane 2 : SMARCA4 knockout HEK-293T cell lysate (20µg)

Lanes 1- 2 : Merged signal (red and green). Green - ab108318 observed at 185 kDa. Red - loading control ab130007 observed at 124 kDa.

ab108318 Recombinant Anti-BRG1 antibody [EPR3912] was shown to specifically 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 and SMARCA4 knockout samples 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] were incubated overnight at 4°C at 1 in 10000 dilution and 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 (<a href='/en-us/products/cell-lines/human-smarca4-brg1-knockout-hek-293t-cell-line-ab255432'>ab255432</a>)

Predicted band size: 185 kDa

Observed band size: 185 kDa

false

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

Lab

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

Lane 1 : Wild-type HEK-293T cell lysate 20 μg
Lane 2 : SMARCA4 knockout HEK-293T cell lysate 20 μg
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 (<a href='/en-us/products/cell-lines/human-smarca4-brg1-knockout-hek-293t-cell-line-ab255432'>ab255432</a>)

Predicted band size: 185 kDa

Observed band size: 185 kDa

false

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (AB263853)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (AB263853)

Allele-2 : 1 bp deletion in exon 4

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (AB263853)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMARCA4 (BRG1) knockout HEK-293T cell lysate (AB263853)

Allele-1 : 7 bp deletion in exon 4

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

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.

style
left-column

Complementary Products

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)

  • 0
  • 0
Cell Lines & Lysates

AB286434

Human SMARCA4 knockout HCT116 cell line

Next Generation Sequencing - Human SMARCA4 knockout HCT116 cell line (AB286434)

  • 0
  • 0
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)

  • 5
  • 10
Proteins & Peptides

AB217407

Recombinant human Vitronectin/S-Protein (Animal Free)

  • 0
  • 0
style
left-column

Reactivity data

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

Product details

Knockout cell lysate achieved by CRISPR/Cas9.

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.

style
left-column

What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab263853-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human SMARCA4 knockout HEK293T cell lysate", "number":"AB263853-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB263853-CMP02", "productcode":"" } ] } } }
style
left-column

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
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
style
left-column
style
left-column

Quality control

STR analysis

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

style
left-column

Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

style
left-column

Product protocols

style
left-column
style
left-column
style
right-column

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