Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Other assay [1]
Submit
Validation data
Reference
Comment
Report error
- Product number
- PA5-28178 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- METTL3 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: 293T, A431, H1299, HeLa, HepG2, Molt-4, Raji.
- Concentration
- 1 mg/mL
Submitted references Inhibition of resistant triple-negative breast cancer cells with low-dose 6-mercaptopurine and 5-azacitidine.
Singh B, Sarli VN, Lucci A
Oncotarget 2021 Mar 30;12(7):626-637
Oncotarget 2021 Mar 30;12(7):626-637
No comments: Submit comment
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using METTL3 Polyclonal Antibody (Product # PA5-28178). Sample (30 µg of whole cell lysate). Lane A: Molt-4. 7.5% SDS PAGE. METTL3 Polyclonal Antibody (Product # PA5-28178) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot using METTL3 Polyclonal Antibody (Product # PA5-28178). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with METTL3 Polyclonal Antibody (Product # PA5-28178) diluted at 1:10,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody, and the signal was developed with Trident ECL plus-Enhanced.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of N6-adenosine-methyltransferase catalytic subunit was performed using 70% confluent log phase MCF7 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with METTL3 Polyclonal Antibody (Product # PA5-28178) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing nucleus and cytoplasm localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification in EVOS™ M7000 Imaging System (Product # AMF7000) and externally deconvoluted (D.Sage et al. / Methods 115 (2017) 28-41).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 1 Validation of selected gene expression data with Western blotting. Lower levels of METTL3, ESRP1, ESRP2, and CD44v and higher levels of CD44s, SNAIL1, and vimentin are seen in SUM149-MA cells as compared to parental cell line. Parental SUM149-Luc cells were cultured in glutamine-containing medium with dialyzed FBS (indicated in the figure as SUM149). SUM149-MA cells (MA) were maintained in a glutamine-free medium with dialyzed FBS for 9 passages and then switched to glutamine-containing medium for 5 passages before preparing cell lysates for this analysis. Filters were re-probed with a beta-actin antibody to normalize sample loading. The beta-actin blot shown here is a re-probe of the CD44 blot. Relative intensities of protein bands, as quantified with the ImageJ software, are shown at the bottom; the values under the CD44 blot are for CD44s.