重组Anti-MARC1抗体[EPR28272-93] (ab317262)

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

Low expression: lung, intestine (PMID: 20861021).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20861021).

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

In Western blot, Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker  (ab186735) staining at 1/2000 dilution.

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

Low expression: lung, heart (PMID: 20861021).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 20861021).

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

In Western blot, Anti-TOMM20 antibody [EPR15581-54] - Mitochondrial Marker  (ab186735) staining at 1/2000 dilution.

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

Exposure time: Lane 1: 26 seconds; Lane 2: 180 seconds

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

This antibody does not cross-react with human MARC2.

In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade  (ab213204) staining at 1/5000 dilution.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling MARC1 with ab317262 at 1/500 (1.006 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human liver. The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling MARC1 with ab317262 at 1/500 (1.006 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hepatocellular carcinoma. The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver (PMID: 20861021). The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Mouse liver cancer tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse liver cancer. The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Rat liver tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat liver. The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Human lung tissue labeling MARC1 with ab317262 at 1/500 (1.006 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue: weak staining on human lung. The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue: no staining on mouse lung (PMID: 20861021). The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue: no staining on mouse cardiac muscle (PMID: 20861021). The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Rat lung tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue: no staining on rat lung. The section was incubated with ab317262 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.

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling MARC1 with ab317262 at 1/1000 (0.503 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue: no staining on rat cardiac muscle. The section was incubated with ab317262 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.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human liver tissue staining MARC1 with ab317262 at a 1:1000 (0.503 ug/ml) dilution, ab202554 anti-COX IV used at 1:500 (0.23 ug/ml) dilution.

Panel A: merged staining of anti-MARC1 (green; Opal™520), anti-COX IV (magenta; Opal™570) on human liver.
Panel B: anti-MARC1 staining in hepatocyte of human liver.
Panel C: anti-COX IV staining in hepatocyte of human liver.
Panel D: nuclear DNA was labeled with DAPI (shown in blue).
Co-staining of MARC1 and COX IV (mitochondrial marker) can be observed.
The section was incubated in two rounds of staining: in the order of ab317262, ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human kidney tissue staining MARC1 with ab317262 at a 1:1000 (0.503 ug/ml) dilution, ab202554 anti-COX IV used at 1:500 (0.23 ug/ml) dilution.

Panel A: merged staining of anti-MARC1 (green; Opal™520), anti-COX IV (magenta; Opal™570) on human kidney.
Panel B: anti-MARC1 staining on several kidney tubules of human kidney.
Panel C: anti-COX IV staining on kidney tubules of human kidney.
Panel D: nuclear DNA was labeled with DAPI (shown in blue).
Co-staining of MARC1 and COX IV (mitochondrial marker) can be observed.
The section was incubated in two rounds of staining: in the order of ab317262, ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining MARC1 with ab317262 at a 1:1000 (0.503 ug/ml) dilution, ab202554 anti-COX IV used at 1:500 (0.23 ug/ml) dilution.

Panel A: merged staining of anti-MARC1 (green; Opal™520), anti-COX IV (magenta; Opal™570) on human colon.
Panel B: anti-MARC1 staining on human colon.
Panel C: anti-COX IV staining on human colon.
Panel D: nuclear DNA was labeled with DAPI (shown in blue).
Co-staining of MARC1 and COX IV (mitochondrial marker) can be observed.
The section was incubated in two rounds of staining: in the order of ab317262, ab202554 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling MARC1 with ab317262 at 1/50 (10.06 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

Confocal image showing mitochondrial staining in HepG2 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab33985 Anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain at 1/1000 (1ug/ml) dilution (Magenta). The 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 (2 ug/ml) dilution.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling MARC1 with ab317262 at 1/50 dilution (1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

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

MARC1 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab317262 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317262 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 2: ab317262 IP in HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab317262 in HepG2 whole cell lysate

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

Exposure time: 119 seconds.