712072

antibody from Invitrogen Antibodies

Targeting: SCARB2 CD36L2, HLGP85, LIMP-2, LIMPII, SR-BII

Antibody data

Submit

Validation data

Product number: 712072 - Provider product page
Provider: Invitrogen Antibodies
Product name: LIMP2 Recombinant Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Other
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.5 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

SCARB2 protein structure - 712072 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of SCARB2 was achieved by transfecting A549 cells with SCARB2 specific validated siRNA (Silencer® select Product # s2651). Western blot analysis (Fig a) was performed using membrane enriched extracts from SCARB2 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blots were probed with LIMP2 Recombinant Rabbit Monoclonal Antibody (Product # 712072, 1:1500 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 1:20,000 dilution). Densitometric analysis of this Western blot is shown in histogram (Fig b). Loss of signal upon siRNA mediated knock down confirms that antibody is specific to SCARB2.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using LIMP2 Recombinant Polyclonal Antibody (Product # 712072) and a 77 kDa band corresponding to SCARB2 was observed across cell lines and tissues tested. Whole cell extracts (20 µg lysate) of A549 (Lane 1), HCT 116 (Lane 2), Hep G2 (Lane 3), MCF7 (Lane 4), OVCAR-3 (Lane 5); and tissue extracts of Mouse Testis (Lane 6) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX), 12 well. 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 (1:1500 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: For immunofluorescence analysis, HepG2 cells were fixed and permeabilized for detection of endogenous SCARB2 using Anti- SCARB2 Recombinant Rabbit Polyclonal Antibody (Product # 712072, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of SCARB2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents lysosomal staining using LysoTracker® Red DND-99 (Product # L7528). Panel d) is a composite image of Panels a, b and c clearly demonstrating co-localization of SCARB2 with LysoTracker which specifically binds to the Lysosomes. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: For immunofluorescence analysis, SH-SY5Y cells were fixed and permeabilized for detection of endogenous SCARB2 using Anti- SCARB2 Recombinant Rabbit Polyclonal Antibody (Product # 712072, 5 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of SCARB2 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating lysosomal localization of SCARB2. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of SCARB2 was achieved by transfecting A549 cells with SCARB2 siRNA (Silencer® select Product # S2652, S2653). Immunofluorescence analysis was performed on untransfected A549 cells (panel a, d), transfected with non-specific scrambled siRNA (panels e, h) and transfected with SCARB2 specific siRNA (panel i, l). Cells were fixed, permeabilized, and labelled with LIMP2 Recombinant Polyclonal Antibody (Product # 712072, 1:100 dilution) followed by Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731, 1:2000 dilution). Nuclei (blue) were stained using ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962), and Alexa Fluor™ Plus 647 Phalloidin (Product # A30107, 1:2000 dilution) was used for cytoskeletal F-actin (red) staining. Absence of specific signal was observed upon siRNA mediated knockdown (panel c, f) confirming specificity of the antibody to SCARB2 (green). The Images were captured at 60X magnification.