702499

antibody from Invitrogen Antibodies

Targeting: STAG2 SA-2, SA2, SCC3B

Antibody data

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Validation data

STAG2 protein structure - 702499 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of STAG2 was achieved by transfecting HeLa cells with STAG2 specific siRNA (Silencer® select Product # s21089). Western blot analysis was performed using nuclear enriched cell lysates from the STAG2 knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti-STAG2 Recombinant Rabbit Monoclonal Antibody (Product # 702499, 1-2 µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Reduction of signal upon siRNA mediated knock down confirms that antibody is specific to STAG2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), HeLa (Lane 2), A-431 (Lane 3), PANC-1 (Lane 4) and MOLT4 (Lane 5). The blots were probed with Anti-STAG2 Recombinant Rabbit Monoclonal Antibody (Product # 702499, 1 µg/mL). A 160 kDa band corresponding to STAG2 was observed across the cell lines tested. The blots were detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:5000 dilution). Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with wet transfer method. The membrane was probed with the relevant primary and secondary Antibody following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: For immunofluorescence analysis, HeLa cells were fixed and permeabilized for detection of endogenous STAG2 using Anti- STAG2 Recombinant Rabbit Monoclonal Antibody (Product # 702499, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Detection and localization of STAG2 (green) in the nucleus and cytoplasm can be clearly observed in interphase cells (a-d). When cell cycle progresses to prophase (e-h) STAG2 can be visualized detaching from chromatin and further progression to metaphase (i-l) and early cytokinesis (m-p) results in complete detachment of STAG2 from chromatin and translocation to cytoplasm.The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of STAG2 was achieved by transfecting HeLa cells with STAG2 specific siRNA (Silencer® select Product # s21089). Immunofluorescence analysis was performed on HeLa cells (untransfected, panel a-d), and cells transfected with STAG2 specific siRNA (panel i-l) or non-specific scrambled siRNA (panels e-h). Cells were fixed, permeabilized, and labelled with Anti-STAG2 Recombinant Rabbit Monoclonal Antibody (Product # 702499, 2 µg/mL), followed by Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Nuclei (blue) were stained using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938), and Rhodamine Phalloidin (Product # R415, 1:300) was used for cytoskeletal F-actin (red) staining. Loss of signal was observed upon siRNA mediated knockdown (panel i-l) confirming specificity of the antibody to STAG2 (green). The images were captured at 60X magnification.