Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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

• IP

Supplier Data

Immunoprecipitation - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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.

All lanes:

Immunoprecipitation - Anti-KPNA2 antibody [EPR25248-95] (<a href='/en-us/products/primary-antibodies/kpna2-antibody-epr25248-95-ab289858'>ab289858</a>)

Predicted band size: 57 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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

All lanes:

Immunoprecipitation - Anti-KPNA2 antibody [EPR25248-95] (<a href='/en-us/products/primary-antibodies/kpna2-antibody-epr25248-95-ab289858'>ab289858</a>)

Predicted band size: 57 kDa

false

• WB

Lab

Western blot - Anti-KPNA2 antibody [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.

All lanes:

Western blot - Anti-KPNA2 antibody [EPR25248-95] (<a href='/en-us/products/primary-antibodies/kpna2-antibody-epr25248-95-ab289858'>ab289858</a>) at 1/1000 dilution

Lane 1:

Human Testis cell lysate at 20 µg

Lane 2:

Human Colon cell lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Lanes 1 - 2:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 57 kDa

Observed band size: 55 kDa

false

• WB

Supplier Data

Western blot - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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.

All lanes:

Western blot - Anti-KPNA2 antibody [EPR25248-95] (<a href='/en-us/products/primary-antibodies/kpna2-antibody-epr25248-95-ab289858'>ab289858</a>) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblasts myoblast) whole cell lysate at 20 µg

Lane 3:

143B (human osteosarcoma) 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 1/100000 dilution

Predicted band size: 57 kDa

Observed band size: 58 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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.

All lanes:

Western blot - Anti-KPNA2 antibody [EPR25248-95] (<a href='/en-us/products/primary-antibodies/kpna2-antibody-epr25248-95-ab289858'>ab289858</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma) 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 1/100000 dilution

Predicted band size: 57 kDa

Observed band size: 58 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-KPNA2 antibody [EPR25248-95] - BSA and Azide free (AB289866)

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).

All lanes:

Western blot - Anti-KPNA2 antibody [EPR25248-95] (<a href='/en-us/products/primary-antibodies/kpna2-antibody-epr25248-95-ab289858'>ab289858</a>) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Lane 4:

Rat colon tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 57 kDa

Observed band size: 58 kDa

false