Anti-B MyB antibody [EPR26321-89]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B MyB antibody [EPR26321-89] (AB314862)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling B MyB with ab314862 at 1/50 (10.4 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab314862 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-B MyB antibody [EPR26321-89] (AB314862)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling B MyB with ab314862 at 1/50 (10.4 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in 293T cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B MyB antibody [EPR26321-89] (AB314862)

Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling B MyB with ab314862 at 1/50 (10.4 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human diffuse large B cell lymphoma. The section was incubated with ab314862 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B MyB antibody [EPR26321-89] (AB314862)

Immunohistochemical analysis of paraffin-embedded mouse Burkitt's lymphoma tissue labeling B MyB with ab314862 at 1/50 (10.4 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse Burkitt's lymphoma. The section was incubated with ab314862 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B MyB antibody [EPR26321-89] (AB314862)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling B MyB with ab314862 at 1/50 (10.4 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat colon. The section was incubated with ab314862 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-B MyB antibody [EPR26321-89] (AB314862)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling B MyB with ab314862 at 1/50 (10.4 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse colon. The section was incubated with ab314862 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Lab

Western blot - Anti-B MyB antibody [EPR26321-89] (AB314862)

This product does not work in A549 cell lysate.

To preserve protein degradation, harvested cells were immediately lysed, followed by rapid gel electrophoresis and membrane transfer procedures.

All lanes:

Western blot - Anti-B MyB antibody [EPR26321-89] (ab314862) at 1/1000 dilution

Lane 1:

Fresh 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

Frozen 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

Fresh A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Frozen A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 100 kDa

false

Exposure time: 20s

• WB

Supplier Data

Western blot - Anti-B MyB antibody [EPR26321-89] (AB314862)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200,000 dilution.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-B MyB antibody [EPR26321-89] (ab314862) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting MYBL2 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 100 kDa

false

Exposure time: 158s

• ChIP-seq

Lab

ChIP-sequencing - Anti-B MyB antibody [EPR26321-89] (AB314862)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab314862 [EPR26321-89]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-B MyB antibody [EPR26321-89] (AB314862)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab314862 [EPR26321-89]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-B MyB antibody [EPR26321-89] (AB314862)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10^7 cells and 8 µg of ab314862 [EPR26321-89]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• WB

Supplier Data

Western blot - Anti-B MyB antibody [EPR26321-89] (AB314862)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. This antibody does not cross-react with human MYBL1 and human MYB. All the recombinant proteins were made in-house. In western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) staining at 1/5000 dilution.

All lanes:

Western blot - Anti-B MyB antibody [EPR26321-89] (ab314862) at 1/1000 dilution

Lane 1:

Purified His-tagged human MYBL2 fragment at 20 ng

Lane 2:

Purified His-tagged human MYBL1 recombinant protein at 200 ng

Lane 3:

Purified His-tagged human MYB recombinant protein at 20 ng

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 36 kDa

false

Exposure time: 92s