Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [5]
- Immunocytochemistry [3]
- Immunohistochemistry [3]
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Validation data
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- Product number
- PA5-28777 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- eRF1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- Recommended positive controls: Neuro 2A, C8D30, NIH-3T3, Raw264.7, C2C12, PC-12, Rat2.
- Concentration
- 0.91 mg/mL
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of eRF1 using 30 µg of MOLT4 lysate. Samples were loaded onto a 10% SDS-PAGE gel and probed with an ERF1 polyclonal antibody (Product # PA5-28777) at a dilution of 1:1000.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of eRF1 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a eRF1 Polyclonal Antibody (Product # PA5-28777) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of eRF1 was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a eRF1 Polyclonal Antibody (Product # PA5-28777) at a dilution of 1:500.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of eRF1 was achieved by transfecting HeLa cells with eRF1 specific siRNAs (Silencer® select Product # s4840, s4838 ). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the eRF1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-eRF1 Polyclonal Antibody (Product # PA5-28777, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 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 eRF1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of MOLT-4 (Lane 1), Neuro-2A (Lane 2), NIH/3T3 (Lane 3), RAW (Lane 4), Jurkat (Lane 5), C2C12 (Lane 6), HeLa (Lane 7), A549 (Lane 8) and tissue extract of Mouse Lung (Lane 9). The blot was probed with eRF1 Polyclonal Antibody (Product # PA5-28777, 1:3000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 50 kDa band corresponding to eRF1 was observed across the cell lines and tissue tested.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of eRF1 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: eRF1 Polyclonal Antibody (Product # PA5-28777) diluted at 1:500. Red: phalloidin, a cytoskeleton marker. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Immunocytochemistry-Immunofluorescence analysis of eRF1 was performed in A431 cells fixed in 4% paraformaldehyde at RT for 15 min. Green: eRF1 Polyclonal Antibody (Product # PA5-28777) diluted at 1:500. Red: alpha Tubulin, a cytoskeleton marker. Blue: Hoechst 33342 staining.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of eRF1 was performed using 70% confluent log phase HeLa 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 eRF1 Rabbit Polyclonal Antibody(Product # PA5-28777) at 5 microgram/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 merged image showing Cytoplasmic localization. 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)
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- Experimental details
- eRF1 Polyclonal Antibody detects eRF1 protein at cytoplasm on mouse testis by immunohistochemical analysis. Sample: Paraffin-embedded mouse testis. ERF1 Polyclonal Antibody (Product # PA5-28777) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- eRF1 Polyclonal Antibody detects eRF1 protein at cytoplasm on mouse testis by immunohistochemical analysis. Sample: Paraffin-embedded mouse testis. ERF1 Polyclonal Antibody (Product # PA5-28777) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded SW480 xenograft, using eRF1 (Product # PA5-28777) antibody at 1:100 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.