- MA1-177 - Invitrogen Antibodies BACE1 antibody

Hui L, Soliman ML, Geiger NH, Miller NM, Afghah Z, Lakpa KL, Chen X, Geiger JD
Journal of Alzheimer's disease : JAD 2019;67(1):393-410

Bazzari FH, Abdallah DM, El-Abhar HS
Molecules (Basel, Switzerland) 2019 May 24;24(10)

Dursun E, Gezen-Ak D
PloS one 2017;12(11):e0188605

No comments: Submit comment

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of BACE1 was performed by loading 20 µg of the indicated whole cell lysates and 10 µL PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% Milk/TBST for at least 1 hour at room temperature. BACE1 was detected at 56 kD using a BACE1 mouse monoclonal antibody (Product # MA1-177) at a dilution of 1:1000 in blocking buffer overnight at 4°C on a rocking platform, followed by a Superclonal goat anti-Mouse IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of SH-SY5Y (Lane 1), U-87 MG (Lane 2), HeLa (Lane 3) and NIH/3T3 (Lane 4). The blot was probed with Anti-BACE1 Monoclonal Antibody (Product # MA1-177, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 1:4000 dilution). A 48 kDa band corresponding to BACE1 was observed across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of BACE1 was achieved by transfecting HeLa cells with BACE1 specific siRNAs (Silencer® select Product # s24218). Western blot analysis (Fig. a) was performed using whole cell extracts from the BACE1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with BACE1 Monoclonal Antibody (3C1C3) (Product # MA1-177, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/ml, 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 BACE1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of BACE1 (green) in HeLa cells. The cells were fixed with formalin for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 5% Normal Goat Serum (Product # 31872) for 15 minutes at room temperature. Cells were stained with or without BACE1 monoclonal antibody (Product # MA1-177), at a dilution of 1:50 overnight at 4C, and then incubated with a DyLight 488 goat anti-mouse IgG secondary antibody (Product # 35503) at a dilution of 1:1000 for 30 minutes at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ToxInsight at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of BACE1 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 1% BSA for 1 hour at room temperature. The cells were labeled with BACE1 Monoclonal Antibody (3C1C3) (Product # MA1-177) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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). Panel d represents the merged image showing cytoplasmic localization. The images were captured at 60X magnification.

Supportive validation

MA1-177