Anti-Intestinal FABP/FABP2 Antibody Picoband®

Western blot analysis of FABP2/I-FABP using anti-FABP2/I-FABP antibody (PB9943).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: SW620 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-FABP2/I-FABP antigen affinity purified polyclonal antibody (Catalog # PB9943) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for FABP2/I-FABP at approximately 15 kDa. The expected band size for FABP2/I-FABP is at 15 kDa.

IHC analysis of FABP2/I-FABP using anti-FABP2/I-FABP antibody (PB9943).
FABP2/I-FABP was detected in a paraffin-embedded section of mouse intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-FABP2/I-FABP Antibody (PB9943) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

High fat diet mice physiologic parameters and enrichment of m 6 Am-modified genes in GO terms associated with metabolic processes. a Gain in weight of mice fed a high fat diet (HFD) or regular chow diet across weeks. N = 10 (5 HDF mice, and 5 Chow lean control mice), error bars denote SEM. b NMR measurement of lean and fat mass body composition, indicating that the gain in weight was mainly in fat mass. N = 10 (5 HDF mice, and 5 Chow lean control mice). c Fold change of read end counts (IP/Input) within a sliding window of 5 base-pairs around a reported adenosine TSS in the Fabp2 gene in lean (chow) and HFD mice. All adenosines in the sequence are indicated. d Sequence logo of the m 6 Am peaks around and upstream the annotated TSS within the DNA, portraying the canonical m 6 Am genomic consensus. e . Gene ontology analysis of m 6 Am and CDS m 6 A (>50 nt of TSS) showing a clear enrichment of m 6 Am- but not in m 6 A-modified genes, in GO terms associated with metabolic processes. Source data are provided as a Source Data file.
Index in PubMed under a CC BY license. PMID: 34893620

m 6 Am significantly regulates mRNA and protein levels upon high fat diet. a mRNA expression heatmap of >1.5 fold differentially expressed genes with differential m 6 Am methylation in HFD. Heatmap colors represent differential mRNA expression fold-change levels in the respective gene between mice samples (green - lower expression; red - higher expression). Right panel indicates if m 6 Am was gained (light red - HFD specific) or lost (light green - chow specific) in HFD. b Heatmap quantification of the mRNA differentially expressed genes that gained or lost m 6 Am upon HFD. Two-tailed p -value of the Chi-square test is reported. c Log protein expression levels of m 6 Am or m 6 A methylated genes versus non-methylated genes in lean control (chow) mice. Two-tailed p -values of the Mann Whitney U test are reported, covering 2246, 1998, and 5737 genes with the respective locations. Box plots surround the 1–3 quartiles, whiskers denote 1.5 interquartile range. d Full proteomic profiling proportions of genes with protein differential expression >1.5 fold, which gained or lost m 6 Am upon HFD. Two-tailed p -value of the Chi-square test is reported. Full proteomic abundance profiling was conducted on two HFD mice samples and two lean chow control mice samples covering a total of 5914 proteins detected. e Western blot of Fabp2 and Fabp5 genes, which were found decorated with m 6 Am only in lean mice. The results show a clear repeating pattern of overexpression across all lean biological replicates ( N = 5) versus all fat biological replicates ( N = 4). See Supplementary Fig. for the quantification of these signals. The available molecular weight markers can be seen within the full scan blots which are available as Source data accompanying this figure, provided as a Source Data file.
Index in PubMed under a CC BY license. PMID: 34893620

IHC analysis of FABP2/I-FABP using anti-FABP2/I-FABP antibody (PB9943).
FABP2/I-FABP was detected in a paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-FABP2/I-FABP Antibody (PB9943) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of FABP2/I-FABP using anti-FABP2/I-FABP antibody (PB9943).
FABP2/I-FABP was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-FABP2/I-FABP Antibody (PB9943) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IF analysis of Intestinal FABP using anti-Intestinal FABP antibody (PB9943).
Intestinal FABP was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-Intestinal FABP Antibody (PB9943) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.