Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunocytochemistry [1]
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Validation data
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- Product number
- PA1-12527 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- beta Amyloid Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with rabbit, pig, bovine, guinea pig and chicken based on sequence homology.
- Concentration
- 1 mg/mL
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Supportive validation
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- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-beta Amyloid Polyclonal Antibody (Product # PA1-12527) and a ~120 and 90 kD band 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 MCF-7 (Lane 1), PC-3 (Lane 2), SH-SY5Y (Lane 3), U-87 MG (Lane 4), K-562 (Lane 5), HEL 92.1.7 (Lane 6) and tissue extracts of Mouse Brain (Lane 7), Mouse Skeletal Muscle (Lane 8), Rat Brain (Lane 9) and Rat Skeletal Muscle (Lane 10) 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 (2ug/ml) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L), Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 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
- Immunofluorescence analysis of Amyloid Precursor Protein 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 1 hour at room temperature. The cells were labeled with APP Polyclonal Antibody (Product # PA112527) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) 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.