PA1-16817

antibody from Invitrogen Antibodies

Targeting: NPC1 SLC65A1

Immunoprecipitation

Immunohistochemistry

Immunoelectron microscopy

Antibody data

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Validation data

Product number: PA1-16817 - Provider product page
Provider: Invitrogen Antibodies
Product name: NPC1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: By Western blot, PA1-16817 detects heterogeneously glycosylated NPC1 protein with prominent bands at ~170 and ~220 kDa. Suggested positive control: human fibroblast cell lysate.
Reactivity: Human, Mouse, Rat, Hamster, Porcine
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

NPC1 protein structure - PA1-16817 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot detection of human NPC1 in 20 µg of human fibroblast cell lysate using Product # PA1-16817.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout validation by Western blot analysis of NPC1 in lysates of HeLa human cervical epithelial carcinoma parental cell line and NPC1 knockout (KO) HeLa cell line. Samples were incubated in NPC1 polyclonal antibody (Product # PA1-16817) using a dilution of 1:1000 followed by a HRP-conjugated Anti-Rabbit IgG secondary antibody. Specific band was detected for NPC1 at approximately 240-260 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of NPC intracellular cholesterol transporter 1 was achieved by transfecting HeLa with NPC intracellular cholesterol transporter 1 specific siRNAs (Silencer® select Product # s9668, s9669). Western Blot analysis (Fig. a) was performed using Whole cell extracts from the NPC intracellular cholesterol transporter 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The Blot was probed with NPC1 Polyclonal Antibody (Product # PA1-16817, 1:1000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 NPC intracellular cholesterol transporter 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot was performed using Anti-NPC1 Polyclonal Antibody (Product # PA1-16817) and a 180 kDa band corresponding to NPC intracellular cholesterol transporter 1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of U-87 MG (Lane 1), Hep G2 (Lane 2) 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 (1:1000) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000) using the iBright FL 1000 (Product # A32752).