LAMC2 Recombinant Rabbit Monoclonal Antibody [PSH07-27]

☑ Relative expression (RE)

Western blot analysis of LAMC2 on different lysates with Rabbit anti-LAMC2 antibody (HA722815) at 1/5,000 dilution.

Lane 1: A431 cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (low expression) (20 µg/Lane)
Lane 3: HepG2 cell lysate (low expression) (20 µg/Lane)
Lane 4: HaCaT cell lysate (low expression) (20 µg/Lane)
Lane 5: NCI-H441 cell lysate (20 µg/Lane)
Lane 6: Mouse skin tissue lysate (40 µg/Lane)
Lane 7: Mouse stomach tissue lysate (40 µg/Lane)
Lane 8: Rat skin tissue lysate (40 µg/Lane)
Lane 9: Rat stomach tissue lysate (40 µg/Lane)

Predicted band size: 131 kDa
Observed band size: 100/140 kDa

Exposure time: 2 minutes; 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 (HA722815) at 1/5,000 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 LAMC2 on different lysates with Rabbit anti-LAMC2 antibody (HA722815) at 1/5,000 dilution.

Lane 1: KYSE-150-parental cell lysate
Lane 2: KYSE-150-LAMC2 KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 140/90 kDa
Observed band size: 140/90 kDa

Exposure time: 15 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 (HA722815)) 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.

Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-LAMC2 antibody (HA722815) 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 (HA722815) 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.

Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-LAMC2 antibody (HA722815) 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 (HA722815) 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.

Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-LAMC2 antibody (HA722815) 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 (HA722815) 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.

☑ Relative expression (RE)

Immunocytochemistry analysis of A431 (positive) and A549 (low expression) labeling LAMC2 with Rabbit anti-LAMC2 antibody (HA722815) at 1/100 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-LAMC2 antibody (HA722815) at 1/100 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.

☑ Relative expression (RE)

Flow cytometric analysis of A549 (left, low expression) and A431 (right, positive) cells labeling LAMC2.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722815, 1/5,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

LAMC2 was immunoprecipitated from 0.2 mg A431 cell lysate with HA722815 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722815 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: A431 cell lysate (input)
Lane 2: HA722815 IP in A431 cell lysate
Lane 3: Rabbit IgG instead of HA722815 in A431 cell lysate

Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 2 seconds; ECL: K1801