- MA1-980-A647 - Invitrogen Antibodies MYC antibody

Csizmar CM, Petersburg JR, Hendricks A, Stern LA, Hackel BJ, Wagner CR
Bioconjugate chemistry 2018 Apr 18;29(4):1291-1301

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Myc Epitope Tag was performed by loading various amounts of E. coli lysate containing a multi-epitope tagged protein per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a low fluorescence PVDF membrane and blocked with Sea Block blocking buffer for at least 1 hour. The membrane was probed with a AlexaFluor 647-conjugated Myc Epitope Tag monoclonal antibody (Product # MA1-980-A647) at a dilution of 1:500 for 1 hour at room temperature on a rocking platform and washed in TBS-0.1% Tween-20. Detection was performed using a fluorescence imaging system.
Conjugate: Red dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Myc Tag Polyclonal Antibody (Product # MA1-980-A647) by loading whole cell extracts of untransfected and transiently transfected HEK-293E lysates: untransfected, 40 µg (Lane 1), empty vector control, 60 µg (Lane 2), Myc-p65-V5, 60 µg (Lane 3), Myc-p65-V5, 40 µg (Lane 4), Myc-p65-V5, 20 µg (Lane 5), Myc-p65-V5, 10 µg (Lane 6), p65-HA, 60 µg (Lane 7) and 25 ng of Positope (Lane 8) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). A ~65 kDa band corresponding to Myc-p65-V5 was observed in HEK293E transfected lysates on probing with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Positope (Product # R90050) is a 53 kDa recombinant protein consisting of multiple epitope tags but lacks HA tag, which has been used as a positive control for c-Myc tag detection. No cross-reactivity was seen with HA-tagged p65 expressing lysate. Figure 1a is a non-overlay chemiluminescent image where construct Myc-p65-V5 has picked up 65 kDa band at different lysate concentrations and figure 1b is a fluorescent image of the same.
Conjugate: Red dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of HeLa cells transfected with a construct containing a MYC Epitope Tag. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 inPBS for 15 minutes at room temperature and blocked with 3% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were stained with AlexaFluor 647 conjugated MYC-tag monoclonal antibody (Product # MA1-980-A647) at a dilution of 1:25 for 1 hour at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.
Conjugate: Red dye

Supportive validation

MA1-980-A647