Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [3]
- Other assay [1]
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Validation data
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- Product number
- PA5-17829 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Amyloid Precursor Protein 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
- 197 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis APP was performed by loading 30 µg of THP-1 whole cell lysate per well onto an SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat dry milk in TBST for 1 hour at room temperature. The membrane was probed with an APP polyclonal antibody (Product # PA5-17829) at a dilution of 1:500 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Detection was performed using ECL substrate. Data courtesy of the Innovators Program.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of beta amyloid was performed by loading 30 µg of U87MG wild type (Lane 1), U87MG cas9 control (Lane 2), U87MG - beta amyloid knockout (Lane 3) whole cell extracts. The blot was probed with Anti-beta amyloid Polyclonal Antibody (Product # PA5-17829) (1:1000 dilution) and Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036) (1:4000 dilution). Loss of signal upon CRISPR mediated knockout (KO) confirms that antibody is specific to beta amyloid.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-beta Amyloid Polyclonal Antibody (Product # PA5-17829) and ~~120, 90 kDa bands corresponding to beta Amyloid was observed across the panel tested except for K-562, HEL 92.1.7, Mouse Skeletal Muscle and Rat Skeletal Muscle which are reported to be negative. Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), SH-SY5Y (Lane 2), K-562 (Lane 3) and HEL 92.1.7 (Lane 4) (Fig.a) and tissue extracts (30 ug lysate) of Mouse Brain (Lane 1), Mouse Skeletal Muscle (Lane 2), Rat Brain (Lane 3) and Rat Skeletal Muscle (Lane 4) (Fig.b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). 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 APP (green) HEK293T cells. Cells fixed with 4% formaldehyde were permeabilized and blocked with 1X PBS containing 5% BSA and 0.3% Triton X-100 for 1 hour at room temperature. Cells were probed with an APP polyclonal antibody (Product # PA5-17829) at a dilution of 1:100 overnight at 4°C in 1X PBS containing 1% BSA and 0.3% Triton X-100, washed with 1X PBS, and incubated with a fluorophore-conjugated goat anti-rabbit IgG secondary antibody at a dilution of 1:200 for 1 hour at room temperature. Nuclei (blue) were stained with DAPI. Images were taken on a Leica DM1000 microscope at 40X magnification. Data courtesy of the Innovators Program.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of APP in SH-SY5Y cells using an APP polyclonal antibody (Product # PA5-17829) (A) (red) and compared to an isotype control (B) showing cytoplasmic and ER staining. DNA is labeled using a fluorescent blue dye.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of beta Amyloid was performed using 70% confluent log phase PC-3. 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 1 hour at room temperature. The cells were labeled with beta Amyloid Polyclonal Antibody (Product # PA5-17829) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790) 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 localization in plasma membrane, cytoplasm and nucleus. Panel e represents negative cell line K-562 with no expression. 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)
- Main image
- Experimental details
- Immunoprecipitation of APP was performed on HEK293T cells. Antigen-antibody complexes were formed by incubating 500 µg of HEK293T whole cell lysate (in 500 µL volume) with 5 µL of an APP polyclonal antibody (Product # PA5-17829) overnight at 4°C. The immune complexes were captured on 30 µL of protein G agarose, washed extensively, and eluted with 6X Laemmli buffer. Samples were resolved on an 8% SDS-PAGE gel, transferred to a PVDF membrane, and blocked with 5% milk in TBST for 1 hour at room temperature. The membrane was probed with an APP polyclonal antibody (Product # PA5-17829) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated mouse anti-rabbit light chain specific secondary antibody at a dilution of 1:40,000 for 1 hour at room temperature. Chemiluminescent detection was performed using ECL substrate. Data courtesy of the Innovators Program.