Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
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Validation data
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- Product number
- MA1-980-A555 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- c-Myc Monoclonal Antibody (9E10), Alexa Fluor™ 555
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 9E10
- Vial size
- 50 µL
- Concentration
- 1 mg/mL
- Storage
- 4° C, do not freeze
Submitted references Identification of the interacting partners of a lysosomal membrane protein in living cells by BioID technique.
Nguyen-Tien D, Suzuki T, Kobayashi T, Toyama-Sorimachi N, Dohmae N
STAR protocols 2022 Jun 17;3(2):101263
STAR protocols 2022 Jun 17;3(2):101263
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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 555-conjugated Myc Epitope Tag monoclonal antibody (Product # MA1-980-A555) 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using c-Myc Monoclonal Antibody (9E10), Alexa Fluor 555 (Product # MA1-980-A555) 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 # NP0322BOX). 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 (1:200 dilution) and detected by fluorescence using the iBright FL 1500 (Product # A44241). 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.
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 555 conjugated MYC-tag monoclonal antibody (Product # MA1-980-A555) at a dilution of 1:50 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.