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AB267269

Human SMARCB1 (SNF5) knockout HEK-293T cell line

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SMARCB1 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, 16 bp deletion in exon 2 and 1 bp insertion in exon 2. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
4 Images
Western blot - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)
  • WB

Lab

Western blot - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)

Lanes 1-4 : Merged signal (red and green). Green - ab192864 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab192864 Anti-SNF5/SMARCB1 antibody [EPR12014-77] was shown to specifically react with SNF5/SMARCB1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267269 (knockout cell lysate ab257688) was used. Wild-type and SNF5/SMARCB1 knockout samples were subjected to SDS-PAGE. ab192864 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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-SNF5/SMARCB1 antibody [EPR12014-77] (<a href='/en-us/products/primary-antibodies/snf5-smarcb1-antibody-epr12014-77-ab192864'>ab192864</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

SMARCB1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human SMARCB1 (SNF5) knockout HEK-293T cell line (ab267269)

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 44 kDa

Observed band size: 50 kDa

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Western blot - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)
  • WB

Lab

Western blot - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)

Lanes 1-4 : Merged signal (red and green). Green - ab126734 observed at 50 kDa. Red - loading control ab8245 observed at 36 kDa.

ab126734 Anti-SNF5/SMARCB1 antibody [EPR6966] was shown to specifically react with SNF5/SMARCB1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267269 (knockout cell lysate ab257688) was used. Wild-type and SNF5/SMARCB1 knockout samples were subjected to SDS-PAGE. ab126734 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at 1 in 1000 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-SNF5/SMARCB1 antibody [EPR6966] (<a href='/en-us/products/primary-antibodies/snf5-smarcb1-antibody-epr6966-ab126734'>ab126734</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

SMARCB1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human SMARCB1 (SNF5) knockout HEK-293T cell line (ab267269)

Lane 3:

Daudi cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 44 kDa,84 kDa

Observed band size: 50 kDa,90 kDa

false

Sanger Sequencing - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)

Allele-2 : 1 bp insertion in exon 2.

Sanger Sequencing - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)
  • Sanger seq

Unknown

Sanger Sequencing - Human SMARCB1 (SNF5) knockout HEK-293T cell line (AB267269)

Allele-1 : 16 bp deletion in exon2

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, 16 bp deletion in exon 2 and 1 bp insertion in exon 2

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Complementary Products

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
Proteins & Peptides

AB127275

Recombinant Human SNF5/SMARCB1 protein

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  • 0
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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
SMARCB1
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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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

SNF5/SMARCB1 also known as INI1 or BAF47 is a member of the SWI/SNF chromatin remodeling complex. It acts in modifying chromatin structure to regulate gene expression. SNF5/SMARCB1 possesses a mass of approximately 47 kDa. The protein expresses ubiquitously in various tissues with notable presence in the nucleus. Through its mechanical activity in chromatin remodeling SNF5/SMARCB1 facilitates transcriptional regulation and chromosomal stability.
Biological function summary

SNF5/SMARCB1 acts as an integral component of the SWI/SNF complex. This complex functions in altering nucleosome positioning on DNA impacting gene expression linked to cell cycle regulation differentiation and DNA repair. By working within this complex SNF5/SMARCB1 helps in the activation or repression of specific gene sets based on cellular requirements displaying an essential role in maintaining normal cellular function. The interplay of SNF5/SMARCB1 with other chromatin-modifying enzymes extends its influence across various genetic pathways.

Pathways

The involvement of SNF5/SMARCB1 in the cell cycle and transcription regulation pathways is acknowledged. It participates intimately in the p53 signaling pathway which is critical for cell cycle checkpoints and apoptotic responses. SNF5/SMARCB1 engages with other proteins such as p53 and RB1 coordinating to maintain genomic integrity and proper transcriptional programming. These pathways spotlight the protein's influence in important cell fate decisions.

SNF5/SMARCB1 mutations or deletions associate closely with malignant rhabdoid tumors an aggressive pediatric cancer and schwannomatosis a disorder involving benign nerve sheath tumors. The loss of SNF5/SMARCB1 disrupts the SWI/SNF complex's function triggering unregulated cell proliferation and cancer progression. Within these disorders the SWI/SNF complex itself which contains other subunits like BRG1 and BRM is also impacted highlighting SMARCB1's critical role in tumor suppression.
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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 SMARCB1
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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'.

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