Antibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Immunocytochemistry [3]
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Validation data
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- Product number
- A21972 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- ECH1 Monoclonal Antibody (9D11AF3)
- Antibody type
- Monoclonal
- Antigen
- Other
- Reactivity
- Human, Rat
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- 9D11AF3
- Vial size
- 100 µg
- Concentration
- 1 mg/mL
- Storage
- 4° C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of ECH1 was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ECH1 (9D11AF3) Mouse Monoclonal Antibody (Product # A21972) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of ECH1 was achieved by transfecting HepG2 cells with ECH1 specific siRNA (Silencer® select Cat # s4437). Immunofluorescence analysis was performed on HepG2 cells (untransfected, panel a,d), transfected with non-specific scrambled siRNA (panels b,e) and transfected with ECH1 specific siRNA (panel c,f) Cells were fixed, permeabilized, and labelled with ECH1 Mouse monoclonal Antibody (Product # A21972, 5 µg/mL), followed by Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal was observed upon siRNA mediated knockdown (panel c,f) confirming specificity of the antibody to ECH1(green). The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Human HDFn cells were paraformaldehyde fixed (4%, 20 min) and Triton X-100 permeabilized (0.1%, 15 min). The cells were incubated with the antibody at 4 µg/mL overnight at 4°C. Secondary antibody: (green) Alexa Fluor® 488 goat anti-mouse IgG (H+L) at a 1/1000 dilution for 1h. 10% Goat serum was used as the blocking agent for all blocking steps. DAPI was used to stain the cell nuclei (blue). Target protein locates mainly in mitochondria.