- 720242 - Invitrogen Antibodies FABP1 antibody

Scott C, Stokes R, Cha KM, Clouston A, Eslam M, Metwally M, Swarbrick MM, George J, Gunton JE
PloS one 2019;14(12):e0225332

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), Tissue extracts (30 µg lysate) Mouse Liver (Lane 2) and Rat Liver (Lane 3). The blots were probed with Anti-FABP1 Polyclonal Antibody (Product # 720242, 1-2 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 14 kDa band corresponding to FABP1 was observed. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 12% Bis-Tris gel (Product # NP0342BOX), 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® 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)
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Experimental details: Western blot was performed using Anti-FABP1 Polyclonal Antibody (Product # 720242) and a 14.2kDa band corresponding to Fatty acid-binding protein, liver was observed across in Hep G2, Mouse Liver and Rat liver, but not in the other tested cell lines and tissues. Whole cell extracts (30 µg lysate) of Hep G2 (Fig. A, Lane 1), HeLa (Fig. A, Lane 2), SH-SY5Y (Fig. A, Lane 3), HaCaT (Fig. A, Lane 4), and Tissue extracts (30 µg lysate) of Mouse Liver (Fig. B, Lane 1), Mouse Kidney (Fig. B, Lane 2), Mouse Adipose (Fig. B, Lane 3), Mouse Mammary gland (Fig. B, Lane 4), Rat Liver (Fig. B, Lane 5) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). 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 (1.5 µg/mL) 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). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Flow Cytometry analysis of endogenous FABP1 was performed on 3T3-L1 cells labeled with Anti-FABP1 Rabbit Polyclonal Antibody (Product# 720242, 5 ug/ 1M cells) or with Rabbit isotype control and detected with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, (Alexa Fluor® 488 conjugate, Product# A27034, 0.4 ug/ml, 1:2500) as represented by the red and pink histograms respectively. The purple histogram represents unstained control cells and the green histogram represents no-primary-Antibody control. A representative of 10,000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer (4468770).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 5 A) Volcano plot of gene expression by PCR array. The gene name is included in each case where expression was significantly altered (p

720242