MA1-24966

antibody from Invitrogen Antibodies

Targeting: APP AD1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-24966

Provider

Invitrogen Antibodies

Product name

beta Amyloid Monoclonal Antibody (DE2B4)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

Protein digestion pretreatment of paraffin sections using formic acid for 2 - 3 minutes is required prior to staining with MA1-24966.

Antibody clone number

DE2B4

Concentration

1 mg/mL

References

Submitted references

Beta-amyloid induces apoptosis of neuronal cells by inhibition of the Arg/N-end rule pathway proteolytic activity.

Kechko OI, Petrushanko IY, Brower CS, Adzhubei AA, Moskalev AA, Piatkov KI, Mitkevich VA, Makarov AA
Aging 2019 Aug 24;11(16):6134-6152

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-beta Amyloid Monoclonal Antibody (DE2B4)(Product # MA1-24966) and a 120, 90kDa band corresponding to beta Amyloid was observed in PC-3 and not in K-562 and HEL 92.1.7. Whole cell extracts (30 µg lysate) of PC-3 (Lane 1), K-562 (Lane 2) and HEL 92.1.7 (Lane 3), 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-Mouse 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 beta Amyloid 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 45 minutes at room temperature. The cells were labeled with beta Amyloid Monoclonal Antibody (DE2B4) (Product # MA1-24966) at 1:200 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32723), (1:3000 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). Panel d represents the merged image showing Cytoplasm, Plasma membrane and Nuclear localization. Panel e represents merged image for K-562 cells showing no staining for APP. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 3 Interaction of Abeta 42 , tRNA, and Ate1. Schematic representation of ELISA assay with immobilized anti-Abeta1-17 antibodies ( A ) or anti-Abeta36-42 antibodies ( B ). ( C ) Detection of tRNA and Ate1 ability to interact with C-terminus of Abeta. ( D ) Same as (C) but with N-terminal region of Abeta. OD 450 - optical density measured at 450 nm. Each value is the mean +- SD of at least four independent experiments; *p < 0.04, **p < 0.01, ***p < 0.001.