GRP78 / BIP Mouse Monoclonal Antibody [C9-9]

☑ Cell treatment (CT)

Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (M1506-2) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate (15 µg/Lane)
Lane 2: HepG2 cell lysate (15 µg/Lane)
Lane 3: Human brain tissue lysate (30 µg/Lane)
Lane 4: Mouse brain tissue lysate (30 µg/Lane)
Lane 5: U-87 MG cell lysate (30 µg/Lane)
Lane 6: RAW264.7 cell lysate (30 µg/Lane)
Lane 7: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 µg/Lane)
Lane 8: Mouse liver tissue lysate (30 µg/Lane)
Lane 9: Rat liver tissue lysate (30 µg/Lane)
Lane 10: Rat pancreas tissue lysate (30 µg/Lane)

Predicted band size: 72 kDa
Observed band size: 70/72 kDa

Exposure time: Lane 1-4: 11 seconds; Lane 5-10: 4 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 (M1506-2) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Immunohistochemical analysis of paraffin-embedded human liver tissue with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 (M1506-2) 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 liver tissue with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 (M1506-2) 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 liver tissue with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 (M1506-2) 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 HepG2 cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (M1506-2) 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 Mouse anti-GRP78 / BIP antibody (M1506-2) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.