Anti-SUZ12 antibody [SUZ220A]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [SUZ220A] (AB126577)

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling SUZ12 with ab126577 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human testis.
The section was incubated with ab126577 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.
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 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [SUZ220A] (AB126577)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling SUZ12 with ab126577 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on human tonsil.
The section was incubated with ab126577 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.
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 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [SUZ220A] (AB126577)

Immunohistochemical analysis of paraffin-embedded (A) Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) cell pellet / (B) SUZ12 knockout HAP1 (HO010044) cell pellet labeling SUZ12 with ab126577 at 1/100 dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Nuclear staining on (A) Wild-type HAP1 cell pellet, no staining on (B) SUZ12 knockout HAP1 (HO010044) cell pellet.
The section was incubated with ab126577 for 30 mins at room temperature, followed by anti-mouse IgG1 antibody (ab125913) for 8 mins during the LeicaDS9800 kit staining procedure.
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 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-SUZ12 antibody [SUZ220A] (AB126577)

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

To minimize protein degradation, cells and tissues were lysed immediately after harvest and loaded onto membranes for Western blotting.

All lanes:

Western blot - Anti-SUZ12 antibody [SUZ220A] (ab126577) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 81 kDa

false

Exposure time: 37s

• WB

Unknown

Western blot - Anti-SUZ12 antibody [SUZ220A] (AB126577)

Note : A purified version of the antibody was used at 1/100 (stock solution at 1mg/ml). This image was from a different batch produced using hybridoma method

All lanes:

Western blot - Anti-SUZ12 antibody [SUZ220A] (ab126577) at 1/100 dilution

Lane 1:

Hek-SUZ12-V5 lysate at 30 µg

Lane 2:

Hek-mTIMP2 lysate at 30 µg

Lane 3:

Hela cell line lysate at 200 µg

Lane 4:

Jeko1 cell line lysate at 200 µg

Lane 5:

Human tonsil lysate at 200 µg

Lane 6:

Human brain lysate at 200 µg

Lane 7:

MT-2 cell line lysate at 200 µg

Lane 8:

3T3 Mouse cell line lysate at 200 µg

Lane 9:

U266 cell line lysate at 200 µg

Predicted band size: 83 kDa

Observed band size: 83 kDa

false

• WB

Lab

Western blot - Anti-SUZ12 antibody [SUZ220A] (AB126577)

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

ab181602 was used as a GAPDH loading control at a 1/200000 dilution.

Performed under reducing conditions.
In Western blot, anti-SUZ12 antibody ab126577 was shown to bind specifically to SUZ12. Target of interest was observed at 81kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in SUZ12 knockout cell line (lane 2).

All lanes:

Western blot - Anti-SUZ12 antibody [SUZ220A] (ab126577) at 1/1000 dilution

Lane 1:

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

Lane 2:

SUZ12 knockout HAP1 whole cell lysate at 20 µg

Secondary

All lanes:

Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution

Observed band size: 81 kDa

false

Exposure time: 81s

• WB

Unknown

Western blot - Anti-SUZ12 antibody [SUZ220A] (AB126577)

Effect of SUZ12 RNA interference (siRNA) in Jeko-1 and Z-138 mantle cell lymphoma cell lines treated with two different hairpins against SUZ12 during three days. A control cell line (empty vector) was used as negative control. Expression of SUZ12 was analyzed using ab126577. A decrease of protein expression was observed in siSUZ12 Jeko-1 and Z-138 cell extracts confirming antibody specificity. Band signals were normalized with tubulin as a loading control. This image was from a different batch produced using hybridoma method

All lanes:

Western blot - Anti-SUZ12 antibody [SUZ220A] (ab126577)

Lanes 1 and 4:

Empty vector

Lanes 2 and 5:

shSUZ12.783

Lanes 3 and 6:

shSUZ12.2076

Predicted band size: 83 kDa

Observed band size: 83 kDa

false