Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human astrocytoma tissue labeling Metallothionein with ab325177 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human astrocytoma.
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

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling Metallothionein with ab325177 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in human liver.
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

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized MG-63 (human osteosarcoma fibroblast); Ramos (human Burkitt's lymphoma B lymphocyte) cells labelling Metallothionein with ab325177 at 1/500 (1.016 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic and nuclear staining in MG-63 cell line (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).
Low expression : Ramos.

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 : ab325177 at 1/500 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded A MG-63 (human osteosarcoma fibroblast) cell pellets B Ramos (human Burkitt's lymphoma B lymphocyte) tissue labeling Metallothionein with ab325177 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining in A : MG-63 and no signal in B : Ramos.
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

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Ramos (human Burkitt's lymphoma B lymphocyte, Left) / MG-63 (human osteosarcoma fibroblast, Right) cells labelling Metallothionein with ab325177 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : Ramos.

• WB

Supplier Data

Western blot - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

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

Low expression : Ramos, small intestine (PMID : 155411360), lung, cerebellum.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23870553).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Metallothionein antibody [EPR29104-22] (<a href='/ko/products/primary-antibodies/metallothionein-antibody-epr29104-22-ab325177'>ab325177</a>) at 1/1000 dilution

Lane 1:

MG-63 (human osteosarcoma fibroblast) transfected with scrambled siRNA control whole cell lysate at 50 µg

Lane 2:

MG-63 transfected with siRNA specifically targeting Metallothionein whole cell lysate at 50 µg

Lane 3:

Ramos (human Burkitt's lymphoma B lymphocyte) at 50 µg

Lane 4:

Human liver tissue lysate at 50 µg

Lane 5:

Human small intestine tissue lysate at 50 µg

Lane 6:

Human lung tissue lysate at 50 µg

Lane 7:

Human cerebellum tissue lysate at 50 µg

Secondary

All lanes:

Western blot at 1/2000 dilution

Observed band size: 15-20 kDa,36 kDa

false

Exposure time: 70s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Metallothionein antibody [EPR29104-22] - BSA and Azide free (AB325189)

This data was developed using ab325177, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling Metallothionein with ab325177 at 1/100 (5.08 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : No staining in human lung.
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