PA1-331B

antibody from Invitrogen Antibodies

Targeting: NR1H2 LXR-b, LXRb, NER, NER-I, RIP15, UNR

Antibody data

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Validation data

NR1H2 protein structure - PA1-331B shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig.1) Western blot analysis was performed on nuclear enriched extracts (30 µglysate) of MCF7 (Lane 1), HeLa (Lane 2), Hep G2 (Lane 3), Jurkat (Lane 4) and HEK-293 (Lane 5). Fig2.) Likewise, western blot analysis was performed on tissue extracts (30 µglysate) of Mouse Liver, Mouse Spleen and Mouse Lung. The blots were probed with LXR alpha/beta Rabbit polyclonal Antibody (Product # PA1-331B, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). A 40 kDa band corresponding to LXR beta was observed across the cell lines tested. Apart from this in Fig. 2, 50 kDa corresponding to LXR alpha form is also observed across all tissue extracts tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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 iBlot® 2 Dry Blotting System (Product # IB21001). 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: Knockdown of LXR alpha/beta was achieved by transfecting HeLa cells with LXR alpha/beta specific siRNAs (Silencer® select Product # s14684, s19568). Western blot analysis (Fig a) was performed using whole cell extracts from LXR alpha/beta knock down cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with Anti- LXR alpha/beta Rabbit polyclonal Antibody (Product # PA1-331B, 1µg/mL) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 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 LXR alpha/beta.