Recombinant Anti-RNA Helicase A antibody [EPR13521] (ab183731)

Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was 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.

Nuclear staining on rat testis. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was 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.

Nuclear staining on mouse testis. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was 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.

Nuclear staining on human testis. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of paraffin-embedded Mouse breast carcinoma tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was 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.

Nuclear staining on mouse breast carcinoma. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling RNA Helicase A with ab183731 at 1/1000 (0.11 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). This section was 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.

Nuclear staining on human breast carcinoma. The section was incubated with ab183731 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

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

ab181602 was used as a GAPDH loading control.

Exposure time: Lanes 1-5: 103 seconds, Lane 6: 26 seconds.

Lysates were freshly made and used immediately to minimize protein degradation.

Blocking/Diluting Buffer and concentration: 5% NFDM/TBST

The observed bands are consistent with what are described in PMID: 28337295

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RNA Helicase A with purified ab183731 at 1/20 dilution (5µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). 

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RNA Helicase A with purified ab183731 at 1/50 dilution (2.2 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney carcinoma tissue sections labeling RNA Helicase A with purified ab183731 at 1/400 dilution (0.28 µg/mL). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Intracellular Flow Cytometry analysis of HeLa cells using ab183731 (unpurified) at a 1/10 dilution (red) or a Rabbit monoclonal IgG (negative) (green). Goat anti rabbit IgG (FITC) secondary used at a 1/150 dilution. 

Immunocytochemistry analysis of HeLa cells (fixative -20℃ Acetone) labeling RNA Helicase A with ab183731 (unpurified) at a 1/250 dilution. Goat anti rabbit IgG (Alexa Fluor^® 488) secondary used at a 1/200 diution.

Immunohistochemical analysis of paraffin embedded Human lung adenocarcinoma tissue labeling RNA Helicase A with ab183731 (unpurified) at a 1/100 dilution. Prediluted HRP conjugated Rabbit IgG secondary used. Counterstained with Hematoxylin. Heat mediated antigen retrieval was performed with EDTA buffer pH 9 before commencing with IHC staining protocol.