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LearnPA5-17212
antibody from Invitrogen Antibodies
Targeting: EIF3A
EIF3, eIF3-p170, eIF3-theta, EIF3S10, KIAA0139, TIF32
Western blot
ImmunoprecipitationAntibody data
- Antibody Data
- Antigen structure
- References [0]
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- Validations
- Western blot [2]
- Immunocytochemistry [2]
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- Product number
- PA5-17212 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- eIF3a 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
- 15.2 µg/mL
- Storage
- -20°C
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Supportive validation
- 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), Caco-2 (Lane 4), NIH/3T3 (Lane 5), C2C12 (Lane 6), PC-12 (Lane 7), Jurkat (Lane 8) and RAW 264.7 (Lane 9). The blot was probed with Anti-eIF3a Polyclonal Antibody (Product # PA5-17212, 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 166 kDa band corresponding to eIF3a was observed across the cell lines tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of eIF3a was achieved by transfecting HeLa with eIF3a specific siRNAs (Silencer® select Product # s16491, s16489). Western blot analysis (Fig. a) was performed using whole cell extracts from the eIF3a knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with eIF3a Polyclonal Antibody (Product # PA5-17212, 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 eIF3a.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of eIF3A in HeLa cells using an EIF3A polyclonal antibody (Product # PA5-17212) (green). Actin filaments are labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of eIF3a 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 eIF3a Rabbit Polyclonal Antibody(Product # PA5-17212) at 1:50 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.
