PA5-48108
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 [7]
- Immunocytochemistry [7]
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- Product number
- PA5-48108 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- AIF Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Reconstitute at 0.2 mg/mL in sterile PBS.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µg
- Concentration
- 0.2 mg/mL
- Storage
- -20° C, Avoid Freeze/Thaw Cycles
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis from lysates of MCF-7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, CTLL-2 mouse cytotoxic T cell line, DA3 mouse myeloma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-human/mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Product # PA5-48108) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody. A specific band was detected for AIF at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of AIF in MCF‚7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, CTLL‚2 mouse cytotoxic T cell line, DA3 mouse myeloma cell line, and PC‚12 rat adrenal pheochromocytoma cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 0.1 µg/mL followed by a HRP-conjugated Anti-Rabbit IgG secondary antibody. A specific band was detected for AIF at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout validation by Western blot analysis of AIF in lysates of HEK293T human embryonic kidney parental cell line and AIF knockout HEK293T cell line (KO). Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 0.1 µg/mL followed by a HRP-conjugated Anti-Rabbit IgG secondary antibody. A specific band was detected for AIF at approximately 65 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in knockout HEK293T cell line. GAPDH is shown as a loading control. This experiment was conducted under reducing conditions.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of AIF in MCF‚7 human breast cancer cell line, Jurkat human acute T cell leukemia cell line, CTLL‚2 mouse cytotoxic T cell line, DA3 mouse myeloma cell line, and PC‚12 rat adrenal pheochromocytoma cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 0.1 µg/mL followed by a HRP-conjugated Anti-Rabbit IgG secondary antibody. A specific band was detected for AIF at approximately 67 kDa (as indicated). This experiment was conducted under reducing conditions.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of AIF in 0.2 mg/mL lysates of Jurkat human acute T cell leukemia cell line and MCF‚7 human breast cancer cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 1 µg/mL. A specific band was detected for AIF at approximately 69 kDa (as indicated). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of AIF was achieved by transfecting HeLa cells 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 # PA5-48108, 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 # PA5-48108) and a 60kDa band corresponding to AIF was observed across cell lines and tissues tested. Whole cell extracts (30ug 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), Mouse Lung (Lane 8) and Rat Kidney (Lane 9) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2002) 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
- Immunocytochemical analysis of Apoptosis Inducing Factor (AIF) was detected in immersion fixed MCF-7 human breast cancer cell line using Rabbit Anti-human/mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Product # PA5-48108) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the 493-conjugated Anti-Rabbit IgG Secondary Antibody (gree and counterstained with DAPI (blue).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemical analysis of Apoptosis Inducing Factor (AIF) was detected in immersion fixed MCF-7 human breast cancer cell line using Rabbit Anti-human/mouse/Rat AIF Antigen Affinity-purified Polyclonal Antibody (Product # PA5-48108) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the 493-conjugated Anti-Rabbit IgG Secondary Antibody (gree and counterstained with DAPI (blue).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of AIF in immersion fixed HeLa human cervical epithelial carcinoma cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 15 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red) and counterstained with DAPI (blue). Specific staining was localized to mitochondria.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of AIF in immersion fixed MCF-7 human breast cancer cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 10 µg/mL for 3 hours at room temperature followed by NorthernLights™ 493-conjugated Anti-Rabbit IgG Secondary Antibody (green) and counterstained with DAPI (blue).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of AIF in immersion fixed staurosporine-stimulated Jurkat human acute T cell leukemia cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 10 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (yellow) and counterstained with DAPI (blue).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunocytochemistry analysis of AIF in immersion fixed HeLa human cervical epithelial carcinoma cell line. Samples were incubated in AIF polyclonal antibody (Product # PA5-48108) using a dilution of 15 µg/mL for 3 hours at room temperature followed by NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red) and counterstained with DAPI (blue). Specific staining was localized to mitochondria.
- 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 # PA5-48108) 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), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing mitochondria localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.