Anti-SUZ12 antibody [SUZ220A] - BSA and Azide free

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [SUZ220A] - BSA and Azide free (AB324760)

This data was developed using ab126577, the same antibody clone in a different buffer formulation.

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] - BSA and Azide free (AB324760)

This data was developed using ab126577, the same antibody clone in a different buffer formulation.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SUZ12 antibody [SUZ220A] - BSA and Azide free (AB324760)

This data was developed using ab126577, the same antibody clone in a different buffer formulation.

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

• WB

Lab

Western blot - Anti-SUZ12 antibody [SUZ220A] - BSA and Azide free (AB324760)

This data was developed using ab126577, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/suz12-antibody-suz220a-ab126577'>ab126577</a>) 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

Lab

Western blot - Anti-SUZ12 antibody [SUZ220A] - BSA and Azide free (AB324760)

This data was developed using ab126577, the same antibody clone in a different buffer formulation.

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] (<a href='/en-us/products/primary-antibodies/suz12-antibody-suz220a-ab126577'>ab126577</a>) 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