Myc tag Recombinant Mouse Monoclonal Antibody [A3C8-R-2]
cat.: HA601081
Product Type: Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity: Species independent
Applications: ELISA, WB, IP, IF-Cell
Clonality: Monoclonal
Clone number: A3C8-R-2
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Isotype: IgG1
Immunogen: Synthetic peptide EQKLISEEDL conjugated to keyhole limpet haemocyanin.
Positive control: C-terminal Myc-tag fusion protein lysate, N-terminal Myc-tag fusion protein lysate.
Recommended Dilutions:
  ELISA
  WB
  WB
  IP
  IF-Cell
1:10,000
1:2,000-1:10,000 (C-Myc)
1:200-1:500 (N-Myc)
Use at an assay dependent concentration.
1:1,000
Alternative names: c-myc tag Myc Epitope Tag
Images
HA601081_1.jpg Fig1: Immunocytochemistry analysis of 293T cells labeling Myc tag with Mouse anti-Myc tag antibody (HA601081) 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 (HA601081) 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.
HA601081_2.jpg Fig2: Western blot analysis of Myc tag on different lysates with Mouse anti-Myc tag antibody (HA601081) at different dilutions.

Lane 1/2: C-terminal Myc-tag fusion protein lysate
Lane 3/4: N-terminal Myc-tag fusion protein lysate

Lysates/proteins at 50 ng/Lane.

Observed band size: 34 kDa

Exposure time: 30 seconds;

12% 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 (HA601081) at different dilutions was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.
HA601081_3.jpg Fig3: Myc-tag was immunoprecipitated in 2µg C-terminal Myc-tag fusion protein lysate with HA601081. Western blot was performed from the immunoprecipitate using HA601081 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: HA601081 IP in C-terminal Myc-tag fusion protein lysate.
Lane 3: Mouse IgG instead of HA601081 IP in C-terminal Myc-tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
HA601081_4.jpg Fig4: Myc-tag was immunoprecipitated in 2µg N-terminal Myc-tag fusion protein lysate with HA601081. Western blot was performed from the immunoprecipitate using HA601081 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: HA601081 IP in N-terminal Myc-tag fusion protein lysate.
Lane 3: Mouse IgG instead of HA601081 IP in N-terminal Myc-tag fusion protein lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
HA601081_5.jpg Fig5: Myc tag Antibody (HA601081) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.