ECM Profiling Antibody Sampler Kit

Western blot analysis of MMP-1 on different lysates with Rabbit anti-MMP-1 antibody (ER31211) at 1/1,000 dilution.

Lane 1: SK-Br-3 cell lysate (15 µg/Lane)
Lane 2: MCF7 cell lysate (15 µg/Lane)
Lane 3: Raji cell lysate (15 µg/Lane)
Lane 4: A431 cell lysate (15 µg/Lane)
Lane 5: HeLa cell lysate (15 µg/Lane)
Lane 6: HUVEC cell lysate (15 µg/Lane)
Lane 7: A549 cell lysate lysate (30 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 54 kDa

Exposure time: 2 minutes;
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 (ER31211) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of MMP-9 on different lysates with Rabbit anti-MMP-9 antibody (ET1704-69) at 1/5,000 dilution.

Lane 1: THP-1 whole cell lysate (20 µg/Lane)
Lane 2: THP-1 treated with 200nM TPA for 10 minutes whole cell lysate (20 µg/Lane)
Lane 3: Rat lung tissue lysate (20 µg/Lane)
Lane 4: Rat spleen tissue lysate (20 µg/Lane)

Predicted band size: 78 kDa
Observed band size: 92 kDa

Exposure time: 2 minutes;
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 (ET1704-69) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

Western blot analysis of MMP-13 on different lysates with Rabbit anti-MMP-13 antibody (ET1702-14) at 1/2,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HEK-293 cell lysate (15 µg/Lane)
Lane 3: Jurkat cell lysate (15 µg/Lane)
Lane 4: MDA-MB-231 cell lysate (15 µg/Lane)
Lane 5: SW480 cell lysate (15 µg/Lane)

Predicted band size: 54 kDa
Observed band size: 57 kDa

Exposure time: 1 minute 2 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 (ET1702-14) 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.

Western blot analysis of Fibronectin on different lysates with Rabbit anti-Fibronectin antibody (ET1702-25) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution.

Lane 1: HepG2 cell lysate (20 µg/Lane)
Lane 2: Caco-2 cell lysate (20 µg/Lane)
Lane 3: SK-MEL-28 cell lysate (20 µg/Lane)
Lane 4: Human kidney tissue lysate (20 µg/Lane)

Predicted band size: 272 kDa
Observed band size: 272 kDa

Exposure time: 3 minutes 20 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 (ET1702-25) at 1/2,000 dilution and competitor's antibody at 1/2,000 dilution were 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.

Western blot analysis of LOX on different lysates with Rabbit anti-LOX antibody (ET1706-31) at 1/1,000 dilution.

Lane 1: PC-3M cell lysate (10 µg/Lane)
Lane 2: Jurkat cell lysate (10 µg/Lane)
Lane 3: Mouse lung tissue lysate (20 µg/Lane)
Lane 4: Rat lung tissue lysate (20 µg/Lane)
Lane 5: Mouse brain tissue lysate (20 µg/Lane)
Lane 6: Rat brain tissue lysate (20 µg/Lane)

Predicted band size: 47 kDa
Observed band size: 47 kDa

Exposure time: 1 minute; 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 (ET1706-31) at 1/1,000 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.

Western blot analysis of COL1A1 on different lysates with Rabbit anti-COL1A1 antibody (HA722517) at 1/1,000 dilution.

Lane 1: HFF-1 cell lysate (20 µg/Lane)
Lane 2: NIH/3T3 cell lysate (20 µg/Lane)
Lane 3: Human lung tissue lysate (40 µg/Lane)
Lane 4: Mouse skin tissue lysate (40 µg/Lane)
Lane 5: Rat skin tissue lysate (40 µg/Lane)

Predicted band size: 139 kDa
Observed band size: 200/139 kDa

Exposure time: Lane 1-4: 1 minute; Lane 5: 3 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 (HA722517) at 1/1,000 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.

Western blot analysis of COL11A1 on different lysates with Rabbit anti-COL11A1 antibody (HA721718) at 1/1,000 dilution.

Lane 1: K-562 cell lysate (20 µg/Lane)
Lane 2: Jurkat cell lysate (20 µg/Lane)
Lane 3: PANC-1 cell lysate (20 µg/Lane)
Lane 4: Saos-2 cell lysate (20 µg/Lane)

Predicted band size: 181 kDa
Observed band size: 150 kDa

Exposure time: 3 minutes;
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 (HA721718) at 1/1,000 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.

Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-MMP-9 antibody (ET1704-69) 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 (ET1704-69) 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.

Immunocytochemistry analysis of HFF-1 cells labeling COL1A1 with Rabbit anti-COL1A1 antibody (HA722517) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-COL1A1 antibody (HA722517) at 1/500 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 (M1305-2, 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.

Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-COL1A1 antibody (HA722517) 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 (HA722517) 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.

Immunofluorescence analysis of paraffin-embedded mouse kidney tissue labeling COL1A1 with Rabbit anti-COL1A1 antibody (HA722517) 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722517, green) at 1/200 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).