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AB257191

Human BRD2 knockout HEK-293T cell lysate

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BRD2 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 exon7 and 1 bp insertion in exon7 and 22 bp deletion in exon7.

View Alternative Names

BRD2_HUMAN, Bromodomain containing 2, Bromodomain-containing protein 2, D6S113E, DKFZp686N0336, FLJ31942, FSH, FSRG1, Female sterile homeotic related gene 1, Female sterile homeotic related gene 1, mouse, homolog of, KIAA9001, NAT, O27.1.1, RING3, RING3 PROTEIN, RNF3, Really interesting new gene 3 protein

5 Images
Western blot - Human BRD2 knockout HEK-293T cell lysate (AB257191)
  • WB

Lab

Western blot - Human BRD2 knockout HEK-293T cell lysate (AB257191)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : BRD2 knockout HEK293T cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)

ab139690 was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate ab257191) was used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab139690 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-BRD2 antibody [EPR7642] - ChIP Grade (<a href='/en-us/products/primary-antibodies/brd2-antibody-epr7642-chip-grade-ab139690'>ab139690</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

BRD2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human BRD2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-brd2-knockout-hek-293t-cell-line-ab267265'>ab267265</a>)

Lane 3:

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: 126 kDa,19 kDa,20 kDa,26 kDa,32 kDa,34 kDa,44 kDa,88 kDa

Observed band size: 110 kDa,26 kDa,50 kDa

false

Western blot - Human BRD2 knockout HEK-293T cell lysate (AB257191)
  • WB

Lab

Western blot - Human BRD2 knockout HEK-293T cell lysate (AB257191)

Lane 1 : Wild-type HEK293T cell lysate (20 ug)
Lane 2 : BRD2 knockout HEK293T cell lysate (20 ug)
Lane 3 : HeLa cell lysate (20 ug)

ab243865 was shown to specifically react with BRD2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267265 (knockout cell lysate ab257191) was used. Wild-type and BRD2 knockout samples were subjected to SDS-PAGE. ab243865 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours 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-BRD2 antibody [BL-167-2A2] (<a href='/en-us/products/primary-antibodies/brd2-antibody-bl-167-2a2-ab243865'>ab243865</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

BRD2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human BRD2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-brd2-knockout-hek-293t-cell-line-ab267265'>ab267265</a>)

Lane 3:

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: 88 kDa

Observed band size: 110 kDa

false

Sanger Sequencing - Human BRD2 knockout HEK-293T cell lysate (AB257191)
  • Sanger seq

Unknown

Sanger Sequencing - Human BRD2 knockout HEK-293T cell lysate (AB257191)

Allele-2 : 1 bp deletion in exon7

Sanger Sequencing - Human BRD2 knockout HEK-293T cell lysate (AB257191)
  • Sanger seq

Unknown

Sanger Sequencing - Human BRD2 knockout HEK-293T cell lysate (AB257191)

Allele-3 : 1 bp insertion in exon7

Sanger Sequencing - Human BRD2 knockout HEK-293T cell lysate (AB257191)
  • Sanger seq

Unknown

Sanger Sequencing - Human BRD2 knockout HEK-293T cell lysate (AB257191)

Allele-1 : 22 bp deletion in exon7

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 exon7 and 1 bp insertion in exon7 and 22 bp deletion in exon7.

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Reactivity data

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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, 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
BRD2
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
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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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Product promise

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