ICAM3 Recombinant Rabbit Monoclonal Antibody [PSH20-98] - BSA and Azide free

This data was developed using HA724193, the same antibody clone in a different buffer formulation.
Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (HA724193) at 1/5,000 dilution.

Lane 1: U-937 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: Ramos cell lysate
Lane 4: THP-1 cell lysate
Lane 5: HL-60 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 60 kDa
Observed band size: 100-150 kDa

Exposure time: 17 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA724193) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

☑ Cell treatment (CT)

This data was developed using HA724193, the same antibody clone in a different buffer formulation.
Western blot analysis of ICAM3 on different lysates with Rabbit anti-ICAM3 antibody (HA724193) at 1/5,000 dilution.

Lane 1: Jurkat (Human T-lymphoblastic cells) cell lysate
Lane 2: Jurkat treated with PNGase F cell lysate
Lane 3: U-937 (Human acute monocytic leukemia cells) cell lysate
Lane 4: U-937 treated with PNGase F cell lysate

Lysates/proteins at 10 µg/Lane.
Exposure time: 17 seconds; ECL: K1801

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA724193, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 60 kDa
Observed band size: 100-150/60 kDa

This data was developed using HA724193, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Jurkat cells labeling ICAM3 with Rabbit anti-ICAM3 antibody (HA724193) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ICAM3 antibody (HA724193) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

This data was developed using HA724193, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-ICAM3 antibody (HA724193) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA724193) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

This data was developed using HA724193, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-ICAM3 antibody (HA724193) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA724193) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

This data was developed using HA724193, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-ICAM3 antibody (HA724193) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA724193) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.