Anti-IRAK-1 抗体 [EPR26375-90] (BSA and Azide free) (ab302555)

This data was developed using ab302554, the same antibody clone in a different buffer formulation. 

Blocking and diluting buffer and concentration: Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS. 

The samples were run on a Bis-Tris gel.
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
False colour image of Western blot: Anti-IRAK1 antibody (AB302554) 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, AB302554 was shown to bind specifically to IRAK1. A band was observed at 80 kDa in wild-type HAP1 cell lysates with no signal observed at this size in IRAK1 knockout cell line. To generate this image, wild-type and IRAK1 knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS 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 ab300422, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

This data was developed using ab300422, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

This data was developed using ab302554, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling IRAK-1 with ab302554 at 1/100 dilution (4.53 µg/mL) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining in human colon carcinoma. The section was incubated with ab302554 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

This data was developed using ab302554, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded human lung cancer and adjacent tissue labeling IRAK-1 with ab302554 at 1/100 dilution (4.53 µg/mL) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining in human lung cancer, no staining in the adjacent tissue is observed. The section was incubated with ab302554 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.

This data was developed using ab302554, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded A) Wild-type HAP1 (Human chronic myelogenous leukemia near-haploid cell) and B) IRAK1 knockout HAP1cell pellets labeling IRAK-1 with ab302554 at 1/100 dilution (4.53 µg/mL) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection kit). Positive staining on (A) wild-type HAP1 cell pellet, no staining on (B) IRAK1 knockout HAP1 cell pellet. The section was incubated with ab302554 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 Leica DS9800 (BOND™ Polymer Refine Detection kit).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins was used.