PA1-823A

antibody from Invitrogen Antibodies

Targeting: PPARD FAAR, NR1C2, NUC1, NUCII

Provider product page for PA1-823A

Western blot

Immunocytochemistry

Flow cytometry

Other assay

Antibody data

Antibody Data
Antigen structure
References [24]
Comments [0]
Validations
Western blot [3]
Immunocytochemistry [1]
Flow cytometry [1]
Other assay [15]

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Product number: PA1-823A - Provider product page
Provider: Invitrogen Antibodies
Product name: PPAR delta Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: PA1-823A detects peroxisome proliferator activated receptor (PPAR) delta from rat, human and mouse cells. This antibody does not detect PPAR alpha or PPAR gamma.
Concentration: 1 mg/mL

PPARD protein structure - PA1-823A shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of PPAR delta was performed by loading 25 µg of NIH-3T3 (Lane 1), PC12 (Lane 2), and K562 (Lane 3) cell lysates and a molecular weight protein ladder onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with a blocking buffer at 4ºC overnight. The membrane was probed with a PPAR delta polyclonal antibody (Product # PA1-823A) at a dilution of 1:2000 (NIH-3T3 and K562) and 1:1000 (PC12) overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Results show a band at 49 kDa in all three cell lysates.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis was performed on whole cell extracts (20 µg lysate) of NIH/3T3 (lane 1), THP-1 (lane 2), PC-3 (lane 3), PC-12 (lane 4), SH-SY5Y (lane 5), U 2-OS (lane 6), HCT 116 (lane 7), K562 (lane 8), A-431 (lane 9) and Jurkat (lane 10). The blots were probed with Anti-PPAR delta Rabbit Polyclonal Antibody (Product # PA1-823A, 1:250-1:750 dilution) 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 49 kDa band corresponding to PPAR delta was observed across cell lines 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)
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Experimental details: Western Blot was performed using Anti-PPAR delta Polyclonal Antibody (Product # PA1-823A) and a 50 kDa band corresponding to PPAR delta was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) (1%SDS) of THP-1 (Lane 1) and THP-1 treated with PMA (1 nm for 48hrs) (Lane 2) as seen in Fig (a). Tissue extracts of Rat Thyroid (Lane 1), Rat Lung (Lane 2), Rat Liver (Lane 3) and Rat Kidney (Lane 4) as seen in Fig (b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) 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: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Extended Data Figure 6 PPAR-d is the predominant PPAR family member expressed in intestinal progenitors and mediates the effects of HFD a , PPAR-d is the most abundant PPAR family member in ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) based on RNA-seq data. b , Confirmation of PPAR family member mRNA expression levels in ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) by qRT-PCR ( n =5). c , Genes upregulated in HFD ISCs (Lgr5-GFP hi ) versus control ISCs were enriched in PPAR and LXR/RXR motifs. d , GSEA of RNA-seq data identified enrichment of PPAR-d targets in ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) with a HFD. e , Confirmation of induced PPAR-d target gene expression in flow sorted ISCs (Lgr5-GFP hi ) and progenitors (Lgr5-GFP low ) by qRT-PCR ( n= 5). All fold changes were significant with P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Extended Data Figure 7 HFD and PPAR-d signaling boost nuclear beta-catenin localization and activity in intestinal progenitors a , b , HFD-derived ISCs ( a , Lgr5-GFP hi ) and progenitors ( b , Lgr5-GFP low ) required less Wnt3a and R-spondin to initiate organoids compared to C ISCs as measured by comparing organoid-formation in complete ENRW media, which includes EGF, Noggin, R-spondin, and Wnt3a, versus EN media, which includes EGF and Noggin but lacks Wnt3a and R-spondin ( n =3). C-derived progenitors, in contrast to HFD-derived progenitors, rarely formed organoids in either ENRW or EN media. c-f , HFD increased nuclear beta-catenin localization in flow sorted ISCs and progenitors from HFD ( c , n =5) and GW-treated ( d , n =4) mice as determined by immunofluorescence (red=DAPI, cyan=non-phosphorylated beta-catenin, CST 8814S). At least 100 cells per sample were quantified. Representative images are shown in ( e , f ). g - i , HFD ( g ) and GW ( h ) treatment increased the numbers of ISCs and progenitors with beta-catenin + nuclei as assessed by immunostains ( n =4 each). Representative images are shown in ( i ); arrowheads indicate representative nuclear beta-catenin in ISCs (red) and progenitors (black). j - l , Association of PPAR-d and beta-catenin in C and HFD derived intestinal crypts as shown by immunoprecipitation (IP) ( n=3 ). For western blot source data, see Supplementary Figure 1 . (Unless otherwise indicated, data reflect mean +- s.d. from n independent experi

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 8 ACOX2 inhibits tumor through the PPARalpha signaling pathway. A The GSEA showed that PPAR ( P = 0.004) and FATTY_ACID_METABOLISM ( P = 0.006) were significantly changed. B The PPI network of ACOX2, PPARA, PPARD, and PPARG was constructed by the String database. C The expression of PPARA, PPARD, and PPARG genes in liver cancer tissues and adjacent noncancerou tissues by using RT-qPCR. D The mRNA expression of PPARA, PPARD, and PPARG was detected by RT-qPCR in HepG2 and SMCC-7721 (NC, the normal HCC cell lines; ACOX2-Ov, overexpression of ACOX2; Sh- ACOX2, knockdown ACOX2). E The protein levels of PPARA, PPARD, and PPARG were detected by Western blot in HepG2 and SMCC-7721 (NC, the normal HCC cell lines; ACOX2-Ov, overexpression of ACOX2; Sh- ACOX2, knockdown ACOX2, * P < 0.05; ** P < 0.01; n.s. not significant). F The Pearson correlation analysis between ACOX2 and PPARA, PPARD, and PPARG by the GEPIA database.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: FIGURE 9 Tempol effects on the activators of mitochondrial biogenesis and oxidative phosphorylation. (A) Western blotting analysis of peroxisome proliferator-activated receptor delta (PPARdelta) and peroxisome proliferator-activated receptor-gamma coactivator-1alpha (PGC-1alpha) in the diaphragm (DIA) muscle from the sedentary mdx mice ( mdx Sed), exercise trained mdx mice ( mdx Ex), and exercise trained mdx mice treated with Tempol ( mdx Ex+T). (B) The graphs show the PPARdelta and PGC-1alpha levels in the crude extracts of DIA from the mdx Sed, mdx Ex, and mdx Ex+T groups. (C) Western blotting analysis of the OXPHOS complexes (complex V, III, IV, II, and I) in the DIA muscle from the mdx Sed, mdx Ex, and mdx Ex+T groups. (D) The graphs show the OXPHOS protein in the crude extracts of DIA from the mdx Sed, mdx Ex, and mdx Ex+T groups. The blots of the proteins (top row) and beta-actin (loading control, bottom row) are shown. The intensities of each band were quantified and normalized to those of the corresponding control. All values expressed as mean +- standard deviation (SD). * P

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 LEF1 regulates lipid metabolism in MYC-expressing cells. a Heatmap of acylcarnitines affected by LEF1 siRNA in myc-/- + MYC fibroblast, generated by metabolomics analyses of cells transfected with either control or LEF1 siRNA. b Rat1 myc-/- fibroblasts expressing either vector or human MYC were collected and subjected for RNA-seq analysis and expression levels of genes involving lipid metabolism were compared. c Rat1 myc -/- control fibroblast cells and Rat1 myc -/- cells reconstituted with human MYC were transfected with control or LEF1 siRNAs for 3 days and extracted with NP40 lysing buffer for Western blot assays with the indicated antibodies. d HCEC expressing truncated APC were transfected with either control or LEF1 siRNA for 3 days, and then fractionated into nuclear and cytoplasmic fractions for Western blotting with the indicated antibodies. e DLD1 cells transfected with control of LEF1 siRNAs for three days were extracted with NP40 lysis buffer, and the total lysates were subjected to Western blot assays. f ACAD9 and CADS gene expression in colon cancer and adjacent normal tissues deposited in the TCGA database. Blue: downregulated (< 0.7), grey: no change (0.7-1.3), and red: upregulated (> 1.3). n = 41 pairs. g Human colon tumor tissues (T) and their surrounding benign tissues (B) were extracted with NP40 lysis buffer, and the total lysates were subjected to Western blotting with the indicated antibodies. h Model of MYC and LEF1 positive feedback loop in pro

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 7 Direct effects of beta-RA on adipogenesis. ( A ) Percentage of 3TL1 cells after seven days of treatment with 1 mM beta-RA relative to the number of untreated 3TL1 cells. Cells cultured in proliferative conditions. ( B ) Levels of the proteins SKP2, p27, and CYCA2, which were involved in the control of the cell cycle. The 3TL1 cells were treated for seven days with 1 mM beta-RA in proliferative conditions. ( C ) Levels of the proteins SKP2, p27, and CYCA2, which were involved in cell cycle control. The 3TL1 cells were treated for seven days with 1 mM beta-RA in differentiative conditions. ( D , E ) Oil Red O staining in 3TL1 cells cultured under proliferative ( D ) and proliferative + differentiative ( F ) conditions. The 3TL1 cells were treated with 1 mM beta-RA from day 0 in both conditions and the stains were performed on three different days (2, 4, and 7). ( F ) Percentage of the area corresponding to the Oil Red O stains in the 3TL1 cells in differentiative conditions after days 4 and 7 of treatment with 1 mM beta-RA. ( G , H ) Levels of PPARgamma and PPARdelta in the 3TL1 cells cultured in proliferative + differentiative ( F ) conditions and treated with 1 mM beta-RA. The results in non-differentiated cells are shown in line one as the negative control. ( I , J ) Levels of CoQ 9 in the 3TL1 cells cultured in proliferative conditions ( I ) and differentiative conditions ( J ) and treated with 1 mM beta-RA. Data are expressed as mean +- SD. * p < 0.05, *** p < 0.0

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 3 m 1 A58 methylation in tRNA regulates PPARdelta translation. a Scheme of tRNA m 1 A-seq. b Comparison of misincorporation signals caused by m 1 A58 in tRNAs between liver CSC and non-CSC cells. Red dots represent tRNAs with elevated m 1 A58 methylation levels in CSCs. c Comparison of translation efficiency (TE) between liver CSC and non-CSC cells. Red dots represent genes with increased TE, while black dots represent genes with decreased TE in liver CSCs. d Frequency of codons corresponding to m 1 A58 elevated tRNAs was higher in the genes with increased translation efficiency (TE) in CSCs. e Unbiased pathway analysis of TE upregulated genes in liver CSCs compared with non-CSCs. f Western blotting analysis of PPARdelta expression in indicated CSCs. n = 4 biologically independent samples. g Effect of elevated m 1 A58 methylation levels in tRNA Ala(AGC) and tRNA Glu(CTC) on protein synthesis. Reporter vectors were transfected into Mock overexpressed PLC/PRF/5 cells, WT-TRMT6/TRMT61A or Mut-TRMT6/TRMT61A overexpressed PLC/PRF/5 cells, respectively. Control vectors were also transfected into these three cell lines to normalize translation differences among these cell lines. The expression level of two synonymous codon control Ala-(GCG) and Glu-(GAA) remained unchanged upon TRMT6/TRMT61A expression level manipulation. n = 6 biologically independent samples. Exact P values from left to right: 0.00021, 0.27, 0.45, 0.00086, 0.21, 0.15. h Representative immunohistochemistry ima

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 6 Trmt6/Trmt61a knockout inhibits mouse cholesterol synthesis through Ppardelta pathway. a Macroscopic appearance of livers from Trmt6 f/+ /Trmt61a f/+ ; Alb-Cre (WT) and Trmt6/Trmt61a DKO ( Trmt6 f/f /Trmt61a f/f ; Alb-Cre) mice under DEN-induced cancer or in Myc OE mouse model of HCC. Black asterisks indicate liver tumors. Scale bar, 1 cm. b Quantitation of liver tumor numbers of Trmt6 KO, Trmt61a KO, and Trmt6/Trmt61a DKO in DEN-treated and in Myc OE mice. Results are shown as means +- SD ( n = 10 biologically independent mice). Exact P values from left to right: 0.0075, 0.041, 0.032, 0.0037, 0.045, 0.014. c Representative liver H&E staining and immunohistochemistry images of m 1 A, Ppardelta, and Hmgcs2 (brown) from Trmt6 f/ + /Trmt61a f/+ (WT) and Trmt6 -/- /Trmt61a -/- (DKO) in Myc OE and DEN-treated mice. Black circle indicates tumor region. Scale bar, 100 mum. n = 3 independent samples. d Levels of cholesterol biogenesis related proteins in 10-week-old male Trmt6/Trmt61a deleted (DKO) in Myc OE mice transduced with AAV vectors encoding GFP control (oeGFP) or Ppardelta (oePpardelta) for 8 days. WT: Trmt6 f/+ /Trmt61a f/+ ; Myc OE mice. n = 5 biologically independent mice. e Total serum cholesterol levels in 10-week-old normal-fed male Trmt6/Trmt61a deleted Myc OE (DKO) mice transduced with AAV vectors encoding GFP control (oeGFP) or Ppardelta (oePpardelta) for 8 days. WT: Trmt6 f/+ /Trmt61a f/+ ; Myc OE mice. Data are means +- SD. n = 5 mice per group. Exact P val

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 7 TRMT6/TRMT61A complex inhibitor thiram suppresses self-renewal of liver CSCs and tumor growth. a Quantitative LC-MS/MS analysis of m 1 A/A in total RNA and tRNA purified from PLC/PRF/5 cells treated with vehicle or drugs at IC50. Statistical results as means +- SD, n = 5 biologically independent samples. Exact P values from left to right: 0.0048, 0.0086, 0.0037, 0.0090. b Oncosphere forming assays of HCC primary cells following thiram or vehicle treatment. Scale bar, 100 mum. Right panel represents statistical results as means +- SD. n = 15 independent experiments. c Representative magnetic resonance imaging (MRI) images of independent experimental cohorts, of day 0 and day 15 of mice enrolled in treatment with vehicle (DMSO), thiram (1.6 mg kg -1 ), GSK3787 (5 mg kg -1 ) or thiram combined with GSK3787. Red circles and asterisk indicate visible tumor area that was used to calculate tumor volumes. Scale bar, 1 cm. Tumor volumes were calculated on the basis of MRI images from mice bearing liver tumor with a matched initial tumor volume (Right panel). Graph shows means +- SD. n = 6 mice per group. Exact P values from left to right: 0.00065, 0.028. d Tumor volumes were calculated from mice treated with vehicle or atorvastatin (Atorva., 10 mg kg -1 ). Graph shows means +- SD. n = 6 mice per group. Exact P value: 0.0073. e Survival curve derived from mice bearing HCC PDC tumors, treated with vehicle ( n = 11), thiram ( n = 12), GSK3787 ( n = 10), or thiram combined with GSK