- PA5-27137 - Invitrogen Antibodies ACSL4 antibody

Submitted references Dipeptidase-1 governs renal inflammation during ischemia reperfusion injury.

Lau A, Rahn JJ, Chappellaz M, Chung H, Benediktsson H, Bihan D, von Mässenhausen A, Linkermann A, Jenne CN, Robbins SM, Senger DL, Lewis IA, Chun J, Muruve DA
Science advances 2022 Feb 4;8(5):eabm0142

Embryonal erythropoiesis and aging exploit ferroptosis.

Zheng H, Jiang L, Tsuduki T, Conrad M, Toyokuni S
Redox biology 2021 Oct 30;48:102175

Dual-process brain mitochondria isolation preserves function and clarifies protein composition.

Noterman MF, Chaubey K, Lin-Rahardja K, Rajadhyaksha AM, Pieper AA, Taylor EB
Proceedings of the National Academy of Sciences of the United States of America 2021 Mar 16;118(11)

Tumor suppressor p53 promotes ferroptosis in oxidative stress conditions independent of modulation of ferroptosis by p21, CDKs, RB, and E2F.

Kuganesan N, Dlamini S, Tillekeratne LMV, Taylor WR
The Journal of biological chemistry 2021 Dec;297(6):101365

GSAP regulates lipid homeostasis and mitochondrial function associated with Alzheimer's disease.

Xu P, Chang JC, Zhou X, Wang W, Bamkole M, Wong E, Bettayeb K, Jiang LL, Huang T, Luo W, Xu H, Nairn AC, Flajolet M, Ip NY, Li YM, Greengard P
The Journal of experimental medicine 2021 Aug 2;218(8)

Diet and Exercise Training Influence Skeletal Muscle Long-Chain acyl-CoA Synthetases.

Stierwalt HD, Ehrlicher SE, Robinson MM, Newsom SA
Medicine and science in sports and exercise 2020 Mar;52(3):569-576

Tissue-Specific Ablation of ACSL4 Results in Disturbed Steroidogenesis.

Wang W, Hao X, Han L, Yan Z, Shen WJ, Dong D, Hasbargen K, Bittner S, Cortez Y, Greenberg AS, Azhar S, Kraemer FB
Endocrinology 2019 Nov 1;160(11):2517-2528

Control of antiviral innate immune response by protein geranylgeranylation.

Yang S, Harding AT, Sweeney C, Miao D, Swan G, Zhou C, Jiang Z, Fitzgerald KA, Hammer G, Bergo MO, Kroh HK, Lacy DB, Sun C, Glogauer M, Que LG, Heaton NS, Wang D
Science advances 2019 May;5(5):eaav7999

BMP4 Upregulation Is Associated with Acquired Drug Resistance and Fatty Acid Metabolism in EGFR-Mutant Non-Small-Cell Lung Cancer Cells.

Bach DH, Luu TT, Kim D, An YJ, Park S, Park HJ, Lee SK
Molecular therapy. Nucleic acids 2018 Sep 7;12:817-828

The endogenous subcellular localisations of the long chain fatty acid-activating enzymes ACSL3 and ACSL4 in sarcoma and breast cancer cells.

Radif Y, Ndiaye H, Kalantzi V, Jacobs R, Hall A, Minogue S, Waugh MG
Molecular and cellular biochemistry 2018 Nov;448(1-2):275-286

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of FACL4 using 30 µg of A) HeLa and B) NT2D1 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a FACL4 polyclonal antibody (Product # PA5-27137) at a dilution of 1:1000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of FACL4 using 30 µg of A) NIH-3T3 (B) JC and C) BCL-1 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with a FACL4 polyclonal antibody (Product # PA5-27137) at a dilution of 1:5000.

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 PC-3 (Lane 1), DU 145 (Lane 2), NIH/ 3T3 (Lane 3), HeLa (Lane 4), Hep G2 (Lane 5), Caco-2 (Lane 6) and HT-29 (Lane 7). The blot was probed with Anti-ACSL4 Polyclonal Antibody (Product # PA5-27137, 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 55 kDa band corresponding to ACSL4 was detected across the cell lines tested.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of ACSL4 was performed by separating 30 µg of various whole cell extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a ACSL4 Polyclonal Antibody (Product # PA5-27137) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using ACSL4 Polyclonal Antibody (Product # PA5-27137). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with ACSL4 Polyclonal Antibody (Product # PA5-27137) 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 ACSL4 Polyclonal Antibody (Product # PA5-27137). Various whole cell extracts (30 µg) were separated by 7.5% SDS-PAGE, and the membrane was blotted with ACSL4 Polyclonal Antibody (Product # PA5-27137) 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: Knockdown of ACSL4 was achieved by transfecting HeLa cells with ACSL4 specific siRNAs (Silencer® select Product # s5002, s5000). Western blot analysis (Fig. a) was performed using whole cell extracts from the ACSL4 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blots were probed with ACSL4 Polyclonal Antibody (Product # PA5-27137, 1:1000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/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 ACSL4.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of FACL4 in methanol-fixed HeLa cells using a FACL4 polyclonal antibody (Product # PA5-27137) (Green) at a 1:500 dilution. Alpha-tubulin filaments were labeled with Product # PA5-29281 (Red) at a 1:2000.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunocytochemistry-Immunofluorescence analysis of ACSL4 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: ACSL4 Polyclonal Antibody (Product # PA5-27137) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ACSL4 Polyclonal Antibody detects FACL4 protein at cytoplasm by immunofluorescent analysis. Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min. Green: FACL4 stained by ACSL4 Polyclonal Antibody (Product # PA5-27137) diluted at 1:500. Blue: Fluoroshield with DAPI . Scale bar= 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: ACSL4 Polyclonal Antibody detects FACL4 protein at cell membrane and cytoplasm by immunohistochemical analysis. Sample: Paraffin-embedded mouse kidney. FACL4 stained by ACSL4 Polyclonal Antibody (Product # PA5-27137) diluted at 1:500. Antigen Retrieval: Citrate buffer, pH 6.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: ACSL4 Polyclonal Antibody detects FACL4 protein at cytosol on mouse lymph node by immunohistochemical analysis. Sample: Paraffin-embedded mouse lymph node. ACSL4 Polyclonal Antibody (Product # PA5-27137) dilution: 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: ACSL4 Polyclonal Antibody detects FACL4 protein at cytosol on rat middle brain by immunohistochemical analysis. Sample: Paraffin-embedded rat middle brain. ACSL4 Polyclonal Antibody (Product # PA5-27137) dilution: 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: Fig. 5 Equilibrium distributions of organelle marker proteins and ACSL4 in MCF-7 density gradient fractions. a Equal volume samples from each MCF-7 sucrose gradient fraction were subjected to SDS-PAGE separation and immunoblotted for ACSL4 using two different commercially available antibodies; antibody#1 was supplied by GeneTex and antibody#2 from Invitrogen. The gradient distributions of a panel of organelle marker proteins was similarly determined by immunoblotting. Data presented are representitive of experiments repeated 2-3 times with similar results. b The normalised distributions of anti-ACSL3 and anti-ACSL4 immunoreactivties in the gradient fractions. Western blotting signals were quantified using imageJ software. Data are representitive of experiments repeated 3-4 times with similar results

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 BMP4 Knockdown Influences Metabolism and p53 (A) Heatmap showing and comparing top enriched terms. Enrichment test based on the gene ontology (GO, http://geneontology.org/ ) database was conducted using the significant gene list. Significant enrichments are displayed in blue (p value = 0.0001). (B) Heatmap representing changes in expression of top upregulated and downregulated genes in PC9-Gef cells transfected with control or BMP4 #2 siRNA. (C) PC9-Gef and H1993-Gef cells were transfected with control or BMP4 siRNA for 48 hr, then cell lysates were subjected to real-time PCR (top panels) or immunoblotting (bottom panels). (D) PC9-Gef and H1993-Gef cells were transfected with control or ACSL4 siRNA for 48 hr, then cell lysates were subjected to real-time PCR (top panels) or immunoblotting (bottom panels). (E) Schematic diagram illustrating the proposed BMP4 pathway modulating energy metabolism through ACSL4 and triglycerides. (F) PC9-Gef and H1993-Gef cells were transfected with control and BMP4 #2 siRNA for 48 hr, and then cell lysates were further processed for metabolic analyses as described in the Materials and Methods . (G) PC9-Gef cells were transfected with control or siBMP4 #2 siRNA for 48 hr, and cell lysates were subjected to the phosphor-kinase array. p-p53 (Ser15) expression levels are indicated. (H) Indicated cells were transfected with either siBMP4 #2 or siCTL for 48 hr, and then lysates were analyzed for p-p53 and total p53 by immunoblotting.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of FACL4 was performed in HeLa whole cell extracts using 5 µg of ACSL4 Polyclonal Antibody (Product # PA5-27137). Samples were transferred to a membrane and probed with ACSL4 Polyclonal Antibody as a primary antibody and an HRP-conjugated HRP-conjugated anti rabbit IgG was used as a secondary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1. Ca v 1.2alpha1 fractionates with OMMs and MAMs in semipure mitochondria. ( A ) Representative immunoblots of submitochondrial fractions before and after trypsin digest in semipure mitochondria isolated from WT (+) and brain-Cacna1c brain-KO (-) mice. Subcellular compartments are color coded as ER (blue), MAM-enriched (pink), OMM (light gray), IMM (black), and mitochondrial matrix (dark gray). Trypsin was used to digest synaptic and nonsynaptic mitochondrially enriched fractions; 1% Triton-X was used as a control to solubilize all proteins, rendering them accessible to trypsin digest ( n = 4, combined from three experiments). ( B ) Representative immunoblots from French press disrupted mitochondria on nondisrupted mitochondria (semipure mito), mitoplast-enriched fractions, and OMM-enriched cell fractions ( n = 8, combined from three experiments).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 Identification of HNEJ-1 as a ferroptosis-specific antibody among five monoclonal anti-HNE antibodies. (A) Human fibrosarcoma HT1080 cells were treated with erastin (10 muM) in the presence or absence of ferrostatin-1 (Fer-1, 5 muM) for 12 h. Then, they were harvested and fixed with 4% paraformaldehyde (PFA) for cell block. Immunohistochemistry was performed with HNEJ-1~5, respectively (scale bar = 100 mum). (B) Human fibrosarcoma HT1080 cells were treated with erastin (10 muM) in the presence or absence of Fer-1 (5 muM) or deferoxamine mesylate (DFO, 500 muM) for 12 h. Then, they were fixed with 4% PFA. After blocking, cells were incubated with HNEJ-1 or anti-ACSL4 and finally observed with confocal microscopy (scale bar = 10 mum). (C) Human fibrosarcoma HT1080 cells were treated with RSL3 (0.25 muM) in the presence or absence of Fer-1 (5 muM) or DFO (500 muM) for 3 h. Then, they were fixed with 4% PFA. After blocking, cells were then incubated with HNEJ-1 or anti-ACSL4 antibody and finally observed with confocal microscopy (scale bar = 10 mum). Representative data are shown based on 3 independent experiments and the analysis is shown as means +- SEM (n = 3); ***P < 0.001 vs control (CTRL) unless indicated by bar. Refer to text for details. Fig. 1

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 7. DPEP1-mediated leukocyte recruitment and tubular injury. ( A ) Immunoblotting for LRP2, GPX4, ACSL4, and DPEP1 expression in whole-kidney tissue from Dpep1 -/- and Dpep1 +/+ mice 48 hours after renal IRI or sham operation. ( B ) Kidney IVM in LysM gfp/gfp mice at 90 min following LPS administration with or without cilastatin or LSALT peptide treatment. Scale bars, 100 mum. ( C ) Stationary GFP + leukocytes/field in the kidney were quantified (versus LPS: cilastatin, * P = 0.01; LSALT, ** P = 0.002; n = 4 to 5 per group, ANOVA with Dunnett's post hoc test). ( D ) Kidney IVM in Dpep1 +/+ and Dpep1 -/- mice at 90 min following LPS administration. Labels: leukocytes (CD11b, red), capillaries (QTracker, blue), and tubules (autofluorescence, yellow-green). Scale bars, 100 mum. ( E ) Kidney IVM with SYTOX Red staining in LysM gfp/gfp mice 2 hours following LPS administration. Non-LPS (NT)-treated mice are shown as a control. Labels: leukocytes (LysM-GFP, bright green/yellow), tubules (autofluorescence, dark green), capillaries (QTracker, blue), and necrotic cells (SYTOX, red). Scale bars, 100 mum.

PA5-27137

Antibody data