Myc tag Recombinant Mouse Monoclonal Antibody [A3C8-R-2] - BSA and Azide free

Western blot analysis of Myc tag on different lysates with Mouse anti-Myc tag antibody (HA610018) at 1/100,000 dilution.

Lane 1: 293T transfected with Myc-tagged empty control cell lysate
Lane 2: 293T transfected with Myc-tagged Claudin18.2 (C-terminal) cell lysate
Lane 3: 293T transfected with Myc-tagged Histone H3.1 (N-terminal) cell lysate

Lysates/proteins at 10 µg/Lane.

Exposure time: 1 minute 2 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 (HA610018) at 1/100,000 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.

Immunocytochemistry analysis of 293T cells labeling Myc tag with Mouse anti-Myc tag antibody (HA610018) at 1/1,000 dilution.

293T cells, transfected with Myc-tagged empty control, Claudin18.2 (C-terminal) or Histone H3.1 (N-terminal) expression vector, respectively, were fixed in 4% paraformaldehyde for 10 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-Myc tag antibody (HA610018) at 1/1,000 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.

Myc Tag (R1208-1, red) was stained at 1/200 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

Myc-tag was immunoprecipitated in 2µg C-terminal Myc-tag fusion protein lysate with HA610018. Western blot was performed from the immunoprecipitate using HA610018 at 1/2,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1:5,000 dilution was used for 60 mins at room temperature.

Lane 1: C-terminal Myc-tag fusion protein lysate (input).
Lane 2: HA610018 IP in C-terminal Myc-tag fusion protein lysate.
Lane 3: Mouse IgG instead of HA610018 IP in C-terminal Myc-tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.

Myc-tag was immunoprecipitated in 2µg N-terminal Myc-tag fusion protein lysate with HA610018. Western blot was performed from the immunoprecipitate using HA610018 at 1/500 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1:2,000 dilution was used for 60 mins at room temperature.

Lane 1: N-terminal Myc-tag fusion protein lysate (input).
Lane 2: HA610018 IP in N-terminal Myc-tag fusion protein lysate.
Lane 3: Mouse IgG instead of HA610018 IP in N-terminal Myc-tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.

Myc tag Antibody (HA610018) in indirect ELISA.

Indirect ELISA analysis of Myc tag was performed by coating wells of a 96-well plate with 50 µl per well of Myc tag antigen diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with StartingBlock blocking buffer, and incubated with 50 µl per well of a mouse Myc tag monoclonal antibody starting at a concentration of 20 µg/mL and serially diluting it to a concentration of 1.28 ng/mL for 1 hour at 37℃. The plate was washed and incubated with 50 µl per well of an HRP-conjugated goat anti-Mouse IgG secondary antibody at a dilution of 1:10,000 for 30 minutes at 37℃. Detection was performed using an Ultra TMB Substrate for 5 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.