Anti-NFkB p105 / p50 antibody [EPR25226-156]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling NFkB p105/p50 with ab283688 at 1/4000 (0.134 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human tonsil. The section was incubated with ab283688 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-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labelling NFkB p105/p50 with ab283688 at 1/4000 (0.134 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Cytoplasmic staining on human colon cancer. The section was incubated with ab283688 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-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling NFkB p105/p50 with ab283688 at 1/4000 (0.134 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Negative control : No staining on human liver. The section was incubated with ab283688 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-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

Immunohistochemical analysis of paraffin-embedded Human prostate hyperplasia tissue labelling NFkB p105/p50 with ab283688 at 1/4000 (0.134 ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining on human prostate hyperplasia. The section was incubated with ab283688 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-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

NFκB p105/p50 was immunoprecipitated from 0.35 mg HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate 10 μg with ab283688 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab283688 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate 10 μg

Lane 2 : ab283688 IP in HAP1 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab283688 in HAP1 whole cell lysate

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

Exposure time : 7.75 seconds

All lanes:

Immunoprecipitation - Anti-NFkB p105 / p50 antibody [EPR25226-156] (ab283688)

Predicted band size: 105 kDa

Observed band size: 50 kDa

false

• WB

Lab

Western blot - Anti-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

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

Exposure time : 59 seconds

All lanes:

Western blot - Anti-NFkB p105 / p50 antibody [EPR25226-156] (ab283688) at 1/1000 dilution

All lanes:

Human heart tissue lysate at 20 µg

Secondary

All lanes:

VeriBlot for IP secondary antibody(HRP)(<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Predicted band size: 105 kDa

false

• WB

Lab

Western blot - Anti-NFkB p105 / p50 antibody [EPR25226-156] (AB283688)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBSLanes 1 - 3 : Merged signal (red and green). Green - ab283688 observed at 120, 50 kDa. Red - loading control, ab8245, was observed at 36 kDa.

ab283688 was shown to specifically react with NFkB p105 / p50 when NFkB p105 / p50 knockout samples were used. Wild-type and NFkB p105 / p50 knockout samples were subjected to SDS-PAGE.

ab283688 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-NFkB p105 / p50 antibody [EPR25226-156] (ab283688) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

NFkB p105 / p50 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

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

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 105 kDa

Observed band size: 120 kDa,50 kDa

false