NM23 Recombinant Mouse Monoclonal Antibody [13C2-R]

Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (HA601307) at 1/2,000 dilution.

Lane 1: HEK-293 cell lysate (20 µg/Lane)
Lane 2: MCF7 cell lysate (20 µg/Lane)
Lane 3: Jurkat cell lysate (20 µg/Lane)
Lane 4: Raji cell lysate (20 µg/Lane)
Lane 5: HeLa cell lysate (20 µg/Lane)
Lane 6: K-562 cell lysate (20 µg/Lane)
Lane 7: A549 cell lysate (20 µg/Lane)
Lane 8: A431 cell lysate (20 µg/Lane)
Lane 9: HepG2 cell lysate (20 µg/Lane)
Lane 10: NIH/3T3 cell lysate (20 µg/Lane)
Lane 11: C6 cell lysate (20 µg/Lane)
Lane 12: Mouse kidney tissue lysate (40 µg/Lane)
Lane 13: Mouse liver tissue lysate (40 µg/Lane)
Lane 14: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 17 kDa
Observed band size: 17/20 kDa

Exposure time: 1 minute 59 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 (HA601307) at 1/2,000 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 mouse kidney tissue with Mouse anti-NM23 antibody (HA601307) at 1/2,000 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 (HA601307) at 1/2,000 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 HeLa cells labeling NM23 with Mouse anti-NM23 antibody (HA601307) 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-NM23 antibody (HA601307) 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.

Immunocytochemistry analysis of NIH/3T3 cells labeling NM23 with Mouse anti-NM23 antibody (HA601307) 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-NM23 antibody (HA601307) 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.

Immunocytochemistry analysis of C6 cells labeling NM23 with Mouse anti-NM23 antibody (HA601307) 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-NM23 antibody (HA601307) 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.

☑ Knockdown (KD)

Western blot analysis of NM23 on different lysates with Mouse anti-NM23 antibody (HA601307) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si NM23 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 17 kDa
Observed band size: 17/20 kDa

Exposure time: 30 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 (HA601307) at 1/2,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.