Laminin beta 1 Recombinant Rabbit Monoclonal Antibody [JM099-03]

Western blot analysis of Laminin beta 1 on different lysates with Rabbit anti-Laminin beta 1 antibody (ET1703-14) at 1/5,000 dilution.

Lane 1: SW480 cell lysate
Lane 2: NIH/3T3 cell lysate
Lane 3: MEF cell lysate
Lane 4: C6 cell lysate
Lane 5: Mouse stomach tissue lysate
Lane 6: Rat stomach tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 198 kDa
Observed band size: 250 kDa

Exposure time: 20 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 (ET1703-14) 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.

Western blot analysis of Laminin beta 1 on different lysates with Rabbit anti-Laminin beta 1 antibody (ET1703-14) at 1/2,000 dilution.

Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: C6 cell lysate (15 µg/Lane)
Lane 3: Mouse stomach tissue lysate (20 µg/Lane)
Lane 4: Rat heart tissue lysate (20 µg/Lane)

Predicted band size: 198 kDa
Observed band size: 250 kDa

Exposure time: 3 minutes 10 seconds;

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 (ET1703-14) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NPHS2 (ET7107-34, Red), anti-AQP1 (ET1703-34, Green), anti-Laminin beta 1 (ET1703-14, Cyan) and anti-aSMA (ET1607-53, Magenta) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in four rounds of staining: in the order of ET7107-34 (1/1,000 dilution), ET1703-34 (1/5,000 dilution), ET1703-14 (1/1,000 dilution) and ET1607-53 (1/10,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.

Fluorescence multiplex immunohistochemical analysis of mouse kidney (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-Laminin beta 1 (ET1703-14, Yellow), anti-NPHS2 (ET7107-34, Red) and anti-SLC12A1/NKCC2 (HA721906, Violet) on kidney. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of ET1703-14 (1/1,000 dilution), ET7107-34 (1/1,000 dilution) and HA721906 (1/3,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.

Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Laminin beta 1 antibody (ET1703-14) 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 (ET1703-14) 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.

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-Laminin beta 1 antibody (ET1703-14) 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 (ET1703-14) 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.

Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-Laminin beta 1 antibody (ET1703-14) 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 (ET1703-14) 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.