Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue labeling SUZ12 with ab307891 at 1/200 dilution (2.465 μg/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining in human diffuse large B-cell lymphoma (PMID : 20558579). The section was incubated with ab307891 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 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-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling SUZ12 with ab307891 at 1/200 dilution (2.465 μg/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining in human tonsil germinal center (PMID : 20558579). The section was incubated with ab307891 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 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-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labeling SUZ12 with ab307891 at 1/50 dilution (9.86 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/mL) (Green). Confocal image showing nuclear staining in HeLa 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 dilution (2.5 ug/ml) (Red). 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 dilution (2 ug/mL).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SUZ12 KO HAP1 (human chronic myelogenous leukemia near-haploid cell) cells labeling SUZ12 with ab307891 at 1/50 dilution (9.86 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/mL) (Green). Confocal image showing nuclear staining in parental HAP1 cells. 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 dilution (2.5 ug/ml) (Red). 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 dilution (2 ug/mL).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling SUZ12 with ab307891 at 1/50 dilution (9.86 ug/ml) followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/mL) (Green). Confocal image showing nuclear staining in F9 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 dilution (2.5 ug/ml) (Red). 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 dilution (2 ug/mL).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling SUZ12 with ab307891 at 1/50 dilution (1 ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling SUZ12 with ab307891 at 1/1000 dilution (0.493 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining in mouse testis. The section was incubated with ab307891 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 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-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunohistochemical analysis of paraffin-embedded mouse large B-cell lymphoma tissue labeling SUZ12 with ab307891 at 1/1000 dilution (0.493 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining in mosue large B-cell lymphoma. The section was incubated with ab307891 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 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-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling SUZ12 with ab307891 at 1/1000 dilution (0.493 ug/ml) followed by ready to use LeicaDS9800 (Bond® Polymer Refine Detection). Nuclear staining in rat testis. The section was incubated with ab307891 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 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

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Western blot - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Lysates at 20 µg per lane. The samples were run on a Bis-Tris gel. Performed under reducing conditions. False colour image of Western blot : Anti-SUZ12 antibody ( ab307891) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab307891 was shown to bind specifically to SUZ12. A band was observed at 95 kDa in wild-type HAP1 cell lysates with no signal observed at this size in SUZ12 knockout cell line. To generate this image, wild-type and SUZ12 knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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.

Lane 1:

Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 20 µg

Lane 2:

SUZ12 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Predicted band size: 83 kDa

false

• WB

Supplier Data

Western blot - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Blocking/Dilution buffer : 5% NFDM/TBST. Exposure times : Lane 1 and 2 : 3 minutes Lane 3 and 4 : 114 seconds Lane 5-6 : 3 minutes Lysates of lane 5 and 6 were freshly made and used immediately to minimize protein degradation. The blot of lane 5-6 were developed using a high sensitivity ECL substrate. The lower bands in lane 1-4 may be caused by degradation.

All lanes:

Western blot - Anti-SUZ12 antibody [EPR26230-82] (<a href='/ko/products/primary-antibodies/suz12-antibody-epr26230-82-ab307891'>ab307891</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 3:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 4:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

C6 whole cell lysate at 20 µg

Lane 6:

Neuro-2a whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 95 kDa

true

• WB

Supplier Data

Western blot - Anti-SUZ12 antibody [EPR26230-82] - BSA and Azide free (AB307892)

This data was produced using ab307891 the same antibody clone but in a different buffer. Blocking/Dilution buffer : 5% NFDM/TBST. Exposure times : Lane1 and 2 : 81 seconds Lane 3-5 : 3 minutes Lysates were freshly made and used immediately to minimize protein degradation. The blot of lane 3-5 were developed using a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-SUZ12 antibody [EPR26230-82] (<a href='/ko/products/primary-antibodies/suz12-antibody-epr26230-82-ab307891'>ab307891</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 3:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

NCCIT (human pluripotent embryonic carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 95 kDa

false