PA1-12527

antibody from Invitrogen Antibodies

Targeting: APP AD1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA1-12527

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

Western blot

Supportive validation

Submitted by

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).

Immunocytochemistry

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.