- PA5-22061 - Invitrogen Antibodies FASN antibody

Submitted references Antiobesity and antidiabetic effects of the dairy bacterium Propionibacterium freudenreichii MJ2 in high-fat diet-induced obese mice by modulating lipid metabolism.

An M, Park YH, Lim YH
Scientific reports 2021 Jan 28;11(1):2481

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using FASN Polyclonal Antibody (Product # PA5-22061). Sample (50 µg of whole cell lysate). Lane A: Mouse brain. 5% SDS PAGE. FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using FASN Polyclonal Antibody (Product # PA5-22061). Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FASN Polyclonal Antibody detects FASN protein by western blot analysis. A. 30 µg PC-12 whole cell lysate/extract. B. 30 µg Rat2 whole cell lysate/extract.5% SDS-PAGE. FASN Polyclonal Antibody (Product # PA5-22061) dilution: 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: CRISPR-Cas9 mediated genome editing ofFASN (as confirmed by next generation sequencing) was achieved by using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR806223_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Fig (a) Western blot analysis of FASN was performed by loading 30 µg of HepG2 wild type (Lane 1), HepG2 Cas9 (Lane 2) and HepG2 Cas9 cells transduced with FASN Lentiviral sgRNA (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-FASN Polyclonal Antibody (Product # PA5-22061) using 1:1000 dilution and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177 1:5000 dilution).Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). A loss of signal in sgRNA transduced cells using the LentiArray™ CRISPR product line confirms that antibody is specific toFASN (Fig (b)).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using FASN Polyclonal Antibody (Product # PA5-22061). Sample (30 µg of whole cell lysate). A: Molt-4. 7.5% SDS PAGE. FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using FASN Polyclonal Antibody (Product # PA5-22061). Unstimulatd and stimulatd 3T3-L1 whole cell extracts (20 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:500. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody. (The differentiation stimulated medium is composed by basal medium, 10% FBS, 50 µg/mL gentamicin, 1 nM L-glutamin, 500 µM IBMX, 1 µM dexamethasone, 2 µM rosiglitazone and 1 µg/mL insulin.).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Knockdown of FASN was achieved by transfecting HepG2 with FASN specific siRNAs (Silencer® select Product # s5031, s5032). Western blot analysis (Fig. a) was performed using membrane enriched extracts from the FASN knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with FASN Polyclonal Antibody (Product # PA5-22061, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/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 FASN.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of HepG2 (Lane 1), Caco-2 (Lane 2), A549 (Lane 3), HeLa (Lane 4), NIH/3T3 (Lane 5), NTERA-2 (Lane 6), PC-3 (Lane 7), tissue extracts of Mouse Adipose (Lane 8) and Mouse Lung (Lane 9). The blot was probed with FASN Polyclonal Antibody (Product # PA5-22061, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # , A27036, 0.25 µg/ml, 1:4000 dilution). A band at ~273 kDa corresponding to FASN was observed across the cell lines and tissues tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of FASN was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FASN Polyclonal Antibody detects Fatty Acid Synthase protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse brown adipocyte. Fatty Acid Synthase stained by FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FASN Polyclonal Antibody detects Fatty Acid Synthase protein at cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse white adipocyte. Fatty Acid Synthase stained by FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FASN Polyclonal Antibody detects Fatty Acid Synthase protein at cytoplasm on human breast carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human breast carcinoma. FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FASN Polyclonal Antibody detects Fatty Acid Synthase protein at cytoplasm on human colon carcinoma by immunohistochemical analysis. Sample: Paraffin-embedded human colon carcinoma. FASN Polyclonal Antibody (Product # PA5-22061) diluted at 1:500. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded human ovarian carainoma, using Fatty Acid Synthase (Product # PA5-22061) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FASN antibody immunoprecipitates FASN protein in IP experiments. IP Sample: HeLa whole cell lysate/extract A : 30 µg whole cell lysate/extract of FASN protein expressing HeLa cells B : Control with 2.5 µg of pre-immune rabbit IgG C : Immunoprecipitation of FASN by 2.5 µg of FASN antibody (Product # PA5-22061) 5% SDS-PAGE The immunoprecipitated FASN protein was detected by FASN antibody (Product # PA5-22061) diluted at 1:1,000. Anti-rabbit IgG (HRP) was used as a secondary reagent.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 Effects of MJ2 and hkMJ2 on protein expression levels related to adipogenesis and lipid metabolism in eWAT. Total protein was extracted from eWAT in each group, and the relative protein expression levels of the factors related to ( A ) adipogenesis (PPARgamma and C/EBPalpha), ( B ) lipogenesis (FAS, SCD-1 and ACC) and ( C ) lipolysis (ATGL and HSL) were investigated by western blot. Representative images of each protein are shown, and the relative quantified expression levels indicate the mean +- SD. The p values were determined by ANOVA and Tukey's HSD test. The full-length blots are shown in Supplementary Fig. 3 .

PA5-22061

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