- MA1-753 - Invitrogen Antibodies PSEN2 antibody

Diehlmann A, Ida N, Weggen S, Grünberg J, Haass C, Masters CL, Bayer TA, Beyreuther K
Journal of neuroscience research 1999 May 15;56(4):405-19

Diehlmann A, Ida N, Weggen S, Grünberg J, Haass C, Masters CL, Bayer TA, Beyreuther K
Journal of neuroscience research 1999 May 15;56(4):405-19

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Presenilin 2 was performed by loading 25 µg of SH-SY5Y (lane 1), human brain (lane 2) and PC12 (lane 3) onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a Presenilin 2 monoclonal antibody (Product # MA1-753) at a dilution of 1:200 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~23 and 46 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of PSEN2 was achieved by transfecting HeLa with PSEN2 specific siRNAs (Silencer® select Product # s11296, s11295). Western blot analysis (Fig. a) was performed using Whole cell extracts from the PSEN2 knockdown cells (lane 2) and non-targeting scrambled siRNA transfected cells (lane 1). The blot was probed with PSEN2 Monoclonal Antibody (APS 21) (Product # MA1-753, 1:500 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:6000 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 PSEN2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot was performed using Anti-PSEN2 Monoclonal Antibody (APS 21) (Product # MA1-753) and a 19kDa band corresponding to C-terminal fragment of PSEN2 was observed across cell lines tested. Expression was observed to be high in THP-1 when compared to Raji and Jurkat. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), THP-1 (Lane 2), A549 (Lane 3), Raji (Lane 4), Jurkat (Lane 5) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 (1:500 dilution) 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Presenilin 2 using Anti-Presenilin 2 Monoclonal Antibody (APS 21) (Product # MA1-753) shows staining in A2058 Cells. Presenilin 2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Presenilin 2 (Product # MA1-753) at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Presenilin 2 using Anti-Presenilin 2 Monoclonal Antibody (APS 21) (Product # MA1-753) shows staining in Hela Cells. Presenilin 2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Presenilin 2 (Product # MA1-753) at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Presenilin 2 using Anti-Presenilin 2 Monoclonal Antibody (APS 21) (Product # MA1-753) shows staining in NCI-H460 Cells. Presenilin 2 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Presenilin 2 (Product # MA1-753) at a dilution of 1:100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35503, Goat Anti-Mouse). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Human kidney tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Presenilin 2 (Product # MA1-753) or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

MA1-753