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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [5]
- Immunocytochemistry [3]
- Immunohistochemistry [1]
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- Product number
- MA5-16148 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- TFAM Monoclonal Antibody (18G102B2E11)
- Antibody type
- Monoclonal
- Antigen
- Other
- Description
- Suggested positive control: Hela whole cell lysate.
- Reactivity
- Human, Mouse, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 18G102B2E11
- Vial size
- 100 µL
- Concentration
- 1 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Submitted references Mitochondria as target to inhibit proliferation and induce apoptosis of cancer cells: the effects of doxycycline and gemcitabine.
Dijk SN, Protasoni M, Elpidorou M, Kroon AM, Taanman JW
Scientific reports 2020 Mar 9;10(1):4363
Scientific reports 2020 Mar 9;10(1):4363
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TFAM using a monoclonal antibody (Product # MA5-16148).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TFAM in HeLa whole cell lysate. Sample was incubated in TFAM monoclonal antibody (Product # MA5-16148).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of TFAM in 0.5 mg/mL HeLa lysate. Samples were incubated in TFAM monoclonal antibody (Product # MA5-16148). This experiment was performed under reducing conditions using the 12-230 kDa separation system.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of mtTFA was achieved by transfecting HeLa cells with mtTFA specific siRNAs (Silencer® select Product # s14000, s14001). Western blot analysis (Fig. a) was performed membrane enriched extracts from the HeLa knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with AP2M1 Monoclonal Antibody (Product # MA5-16148, 1:1000) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to mtTFA..
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-mtTFA Mouse Monoclonal Antibody (Product # MA5-16148) and a 26 kDa band corresponding to mtTFA was observed in cell lines tested. Membrane enriched extracts (30 µg lysate) of HeLa (Lane 1), T-47D (Lane 2), MDA-MB-231 (Lane 3) and Caco-2 (Lane 4) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of TFAM using a monoclonal antibody (Product # MA5-16148).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of TFAM using a monoclonal antibody (Product # MA5-16148).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of mtTFA was performed using Caco-2 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 1 hour at room temperature. The cells were labeled with mtTFA Mouse Monoclonal Antibody (Product # MA5-16148) at 1:50 dilution in 0.1% BSA and incubated overnight at 4 degree and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723) at a dilution of 1:2000 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). Panel d represents the composite image showing mitochondrial localization of mtTFA. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification..
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of TFAM in human testis. Samples were incubated in TFAM monoclonal antibody (Product # MA5-16148) followed by DAB with hematoxylin counterstain.
