Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- PA5-17422 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- eIF4G Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 7 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of eIF4G in extracts from 293 and Hela cell lines using eIF4G polyclonal antibody (Product # PA5-17422).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of eIF4G was achieved by transfecting Caco-2 with eIF4G specific siRNAs (Silencer® select Product # s4586, s4585). Western blot analysis (Fig. a) was performed using whole cell extracts from the eIF4G knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with eIF4G Polyclonal Antibody (Product # PA5-17422, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 eIF4G.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), Hep G2 (Lane 2), HEK-293 (Lane 3), NIH/3T3 (Lane 4), C2C12 (Lane 5), PANC-1 (Lane 6), MCF7 (Lane 7), A-431 (Lane 8) and Ramos (Lane 9). The blot was probed with Anti-eIF4G Polyclonal Antibody (Product # PA5-17422, 1:2000 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 220 kDa band corresponding to eIF4G was observed across the cell lines tested. Apart from the desired band a 166 kDa band corresponding to eIF4G isoform and a cleaved product of eIF4G around 120 kDa was also detected across the cell lines.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of eIF4G using a polyclonal antibody (Product # PA5-17422).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of eIF4G 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 eIF4G Rabbit Polyclonal Antibody(Product # PA5-17422) at 1:100 dilution 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)
- Main image
- Experimental details
- Immunohistochemical analysis of eIF4G in paraffin-embedded human lung carcinoma using an EIF4G polyclonal antibody (Product # PA5-17422) showing cytoplasmic localization.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of eIF4G in HeLa cells using a eIF4G polyclonal antibody (Product # PA5-17422) (blue) compared to a nonspecific negative control antibody (red).