Anti-KPNA2 抗体 [EPR25248-95] - BSA and Azide free (ab289866)

False colour image of Western blot: Anti-KPNA2 antibody [EPR25248-95] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab289858 was shown to bind specifically to KPNA2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using ab289858, the same antibody clone in a different buffer formulation. 

This data was developed using ab289858, the same antibody clone in a different buffer formulation. 

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

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

This data was developed using ab289858, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon tissue labelling KPNA2 with ab289858 at 1/5000 (0.104 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Sporadic nuclear staining on human colon. The section was incubated with ab289858 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins. 

This data was developed using ab289858, the same antibody clone in a different buffer formulation. 

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

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

This data was developed using ab289858, the same antibody clone in a different buffer formulation. 

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

Exposure time: 5.5 seconds. 

Low expression: colon (PMID: 16818692 ).
An unknown band at ~35 kDa was detected in mouse testis tissue lysate (lane 1). 

This data was developed using ab289858, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human testis tissue labelling KPNA2 with ab289858 at 1/5000 (0.104 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Nucleus staining on human testis. The section was incubated with ab289858 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

This data was developed using ab289858, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labelling KPNA2 with ab289858 at 1/5000 (0.104 µg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) . Sporadic nuclear staining on human colon cancer (PMID:33821218). The section was incubated with ab289858 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

This data was developed using ab289858, the same antibody clone in a different buffer formulation.

KPNA2 was immunoprecipitated from HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab289858 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289858 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 µg

Lane 2: ab289858 IP in HeLa whole cell lysate

Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab289858 in HeLa whole cell lysate

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

Exposure time: 3.25 seconds

The lower band in lane 2 may be caused by degradation.

This data was developed using ab289858, the same antibody clone in a different buffer formulation.

KPNA2 was immunoprecipitated from C2C12 (mouse myoblasts myoblast) whole cell lysate with ab289858 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289858 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: C2C12 (mouse myoblasts myoblast) whole cell lysate 10 µg

Lane 2: ab289858 IP in C2C12 whole cell lysate

Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab289858 in C2C12 whole cell lysate

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

Exposure time: 3.25 seconds