MA5-25354

AGS5, HDDC1, Mg11, MOP-5, SBBI88

Antibody data

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Validation data

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SAMHD1 in HEK293T cells in untransfected (Left lane) and transfected (Right lane) samples using 5 µg per lane. The samples were separated by SDS-PAGE and probed with SAMHD1 (Product # MA5-25354) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SAMHD1 in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with SAMHD1 (Product # MA5-25354) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SAMHD1 in mouse spleen lysate using 10 µg per lane. Samples were probed with SAMHD1 (Product # MA5-25354) monoclonal antibody at a dilution of 1:200.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of SAMHD1 in HepG2, HeLa, SVT2, A549, COS7, Jurkat, MDCK, PC12 and MCF7 cells using 35 µg per lane. Samples were separated by SDS-PAGE and probed with SAMHD1 (Product # MA5-25354) monoclonal antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-SAMHD1 Monoclonal Antibody (OTI1A1) (Product # MA5-25354) and a 72 kDa band corresponding to SAMHD1 was observed in all cell and tissue lysates tested except Jurkat and U-937. Lower expression of SAMHD1 was observed in Mouse Brain as compared to Mouse Lung and Mouse Spleen [10.1016/s0165-2478(00)00276-5]. Modified whole cell extracts (1% SDS) (30 µg lysate) of THP-1 (Lane 1), Caco-2 (Lane 2), Jurkat (Lane 3), U-937 (Lane 4), Mouse Lung (Lane 5), Mouse Spleen (Lane 6), Rat Spleen (Lane 7), Mouse Brain (Lane 8) and Rat Brain (Lane 9) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX). 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 (1:200 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 Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A ~25 kDa band corresponding to tissue IgG was observed in some mouse tissue models.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of SAMHD1 in COS7 cells. Cells were transfected with a plasmid overexpressing SAMHD1 and probed with a SAMHD1 monoclonal antibody (Product # MA5-25354).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of SAMHD1 in COS7 cells. Cells were transfected with a plasmid overexpressing SAMHD1 and probed with a SAMHD1 monoclonal antibody (Product # MA5-25354).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of SAMHD1 was performed using 70% confluent log phase Caco-2 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 SAMHD1 Monoclonal Antibody (OTI1A1) (Product # MA5-25354) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and plasma membrane localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.