PA5-17638
antibody from Invitrogen Antibodies
Targeting: AIFM1
AIF, AUNX1, CMTX4, DFNX5, NAMSD, PDCD8
Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-17638 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AIF 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
- 33 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of AIF was achieved by transfecting HeLa with AIF specific siRNAs (Silencer® select Product # S17442, S17440). Western blot analysis (Fig. a) was performed using Whole cell extracts from the untransfected cells (lane 1), non-targeting scrambled siRNA transfected cells (lane 2) and AIF knockdown cells (lane 3). The blot was probed with AIF Polyclonal Antibody (Product # PA5-17638, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 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 AIF.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-AIF Polyclonal Antibody(Product # PA5-17638) and a 60kDa band corresponding to AIF was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of K-562 (Lane 1), Hep G2 (Lane 2), MCF7 (Lane 3), L6 (Lane 4), RAW 264.7 (Lane 5), HeLa (Lane 6), Mouse Kidney (Lane 7), Rat Kidney (Lane 8), Mouse Lung (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)
- Main image
- Experimental details
- Immunofluorescent analysis of AIF in HeLa cells using an AIF polyclonal antibody (Product # PA5-17638) (green) showing colocalization with mitochondria that have been labeled with a mitochondrial fluorescent red dye. DNA is labeled using a fluorescent blue dye.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of AIF 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 2% BSA for 45 minutes at room temperature. The cells were labeled with AIF Polyclonal Antibody (Product # PA5-17638) at 5 µg in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (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 dilution). Panel d represents the merged image showing mitochondrial 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 AIF in paraffin-embedded human lung carcinoma using an AIF polyclonal antibody (Product # PA5-17638).