Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [2]
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Validation data
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- Product number
- PA5-97731 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- eIF3d Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Reactivity
- Human, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 3.64 mg/mL
- Storage
- -20°C or -80°C if preferred
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western Blot analysis of eIF3d using a eIF3d Polyclonal antibody (Product # PA5-97731) at a concentration of 3.3 µg/mL. Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, HepG2 whole cell lysate, 293T whole cell lysate, Rat liver tissue. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 105 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western Blot analysis of eIF3d using a eIF3d Polyclonal antibody (Product # PA5-97731) at a concentration of 3.3 µg/mL. Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, HepG2 whole cell lysate, 293T whole cell lysate, Rat liver tissue. A secondary Goat polyclonal antibody to rabbit IgG was applied at a 1:50,000 dilution. Observed band size: 105 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
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- Experimental details
- Western blot was performed using Anti-ACTN4 Polyclonal Antibody (Product # PA5-97731) and a 110 kDa band corresponding to Alpha-actinin-4 was observed across all the cell lines and tissue except LNCaP which is reported to be be a low expressing cell line. Whole cell extracts (30 µg lysate) of DU 145 (Lane 1), PC-3 (Lane 2), LNCaP (Lane 3), HeLa (Lane 4), A549 (Lane 5), A-431 (Lane 6), MCF7 (Lane 7) and PANC-1 (Lane 8) and tissue extract (30 µg lysate) of Mouse Skeletal Muscle (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:4000 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)
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- Experimental details
- Immunofluorescent analysis of eIF3d in A549 cells using a eIF3d polyclonal antibody (Product # PA5-97731) at a dilution of 1:66. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated Goat Anti-Rabbit IgG(H+L). Cells were counter-stained with DAPI.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Alpha-actinin-4 was performed using 70% confluent log phase PC-3 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 45 minutes at room temperature. The cells were labeled with ACTN4 Polyclonal Antibody (Product # PA5-97731) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), 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 LNCaP cells having low expression of GSTP1. Panel f 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
- Immunohistochemical analysis of eIF3d in paraffin embedded human breast cancer using a eIF3d polyclonal antibody (Product # PA5-97731) at a dilution of 1:200. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of eIF3d in paraffin embedded human tonsil tissue using a eIF3d polyclonal antibody (Product # PA5-97731) at a dilution of 1:200. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.