Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded :
(A) SH-SY5Y (human neuroblastoma epithelial cell) cell pellet
(B) T-47D (human ductal breast epithelial tumor epithelial cell) cell pellet
(C) DU 145 (human prostate carcinoma epithelial cell) cell pellet
(D) SW480 (human colorectal adenocarcinoma epithelial cell) cell pellet
labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/2000 (0.249 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) SH-SY5Y cell pellet and (B) T-47D cell pellet, weak staining on (C) DU 145 cell pellet and (D) SW480 cell pellet

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/100 (4.98 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil.

The primary antibody was incubated 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) SW480 (human colorectal adenocarcinoma epithelial cell) cells labelling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/1000 (0.498 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear staining in SH-SY5Y cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : SW480.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326070 at 1/1000 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/100 (4.98 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human breast carcinoma.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/100 (4.98 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/100 (4.98 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse cerebrum.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded Mouse pancreas tissue labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/100 (4.98 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse pancreas.

The primary antibody was incubated 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

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/100 (4.98 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse breast cancer.

The primary antibody was incubated 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

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling Zinc finger MIZ domain-containing protein 1 with ab326070 at 1/1000 (10.22 µg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing nuclear staining in mouse primary neural/glia cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection.

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab326070 at 1/1000 dilution, followed by ab150120 at 1/1000 dilution.
-ve control 2 : ab11267 at 1/500 dilution, followed by ab150081 at 1/1000 dilution.

• WB

Lab

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

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

This antibody does not cross-react with overexpressed human ZMIZ2 by western blot.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible. ZMIZ1 can undergo degradation easily.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(150KDa).

All lanes:

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (ab326070) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a myc-His-tag® fresh whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human ZMIZ2 expression vector containing a myc-His-tag® fresh whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human ZMIZ1 expression vector containing a myc-His-tag® fresh 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: 120 kDa,130 kDa,150 kDa,36 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

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

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible. ZMIZ1 can undergo degradation easily.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (ab326070) at 1/1000 dilution

All lanes:

SK-BR-3 (human breast adenocarcinoma epithelial cell) fresh whole cell lysate at 40 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: 120 kDa,130 kDa,36 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

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

Low expression : SW480

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible. ZMIZ1 can undergo degradation easily.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (ab326070) at 1/1000 dilution

Lane 1:

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

Lane 2:

SW480 (human colorectal adenocarcinoma epithelial cell) fresh whole cell lysate at 40 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: 120 kDa,130 kDa,36 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (AB326070)

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

To minimize protein degradation, tissues were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible. ZMIZ1 can undergo degradation easily.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-Zinc finger MIZ domain-containing protein 1 antibody [EPR29807-525] (ab326070) at 1/1000 dilution

Lane 1:

Mouse brain fresh tissue lysate at 20 µg

Lane 2:

Mouse liver fresh tissue 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: 120 kDa,130 kDa,36 kDa

false

Exposure time: 147s