FACL4 Recombinant Rabbit Monoclonal Antibody

货号：

AWA12612

应用：

WB,IHC-P,IF-T,mIHC,FCM

反应性：

Human,Mouse,Rat

来源：

Rabbit

规格 价格 库存
20μL ¥620 1-3个工作日
50μL ¥1250 1-3个工作日
100μL ¥2200 1-3个工作日

产品概述

Product Details

Host Species:

Rabbit

Reactivity:

Human, Mouse, Rat

Molecular Wt:

Predicted MW: 79 kDa
Observed MW: 79 kDa

Clonality:

Monoclonal

Isotype:

IgG

Concentration:

1.086mg/ml

Other Names:

ACS4; ACSL4; ACSL4/FACL4; LACS 4; LACS4; MRX63; MRX68; FACL4

Formulation:

Liquid in PBS containing 50% glycerol, 0.5% BSA and 0.02% sodium azide.

Purification:

Affinity-chromatography

Storage:

Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20°C storage.

Applications

WB 1:1000-1:5000
IHC-P 1:100-1:1000
IF-T 1:100-1:1000
mIHC 1:100-1:1000
FCM 1:50-1:200

Immunogen
Information

Gene Name:

ACSL4

Protein Name:

Long-chain-fatty-acid--CoA ligase 4

Gene ID:

2182 (Human)
50790 (Mouse)
113976 (Rat)

SwissPro:

O60488 (Human)
Q9QUJ7 (Mouse)
O35547 (Rat)

Immunogen
Information

Subcellular Location:

Mitochondrion outer membrane. Peroxisome membrane. Microsome membrane. Endoplasmic reticulum membrane. Cell membrane.

Immunogen:

Recombinant protein within human FACL4. AA range:range: 561-711.

Specificity:

FACL4 Monoclonal Antibody detects endogenous levels of FACL4 protein.

Product images
	Fig : Western blot analysis of FACL4 on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 2 hour at room temperature. The primary antibody (AWA12612,1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG-HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control:* Lane 1: Hep3B2.1-7 cell Lane 2: HepA1-6 cell Lane 3: Raw264.7 cell Lane 4: Mouse kidney Lane 5: Rat kidney Lane 6: 786-O cell Lane 7: ACHN cell Predicted band size:79 kDa Observed band size:79 kDa

	Fig : Western blot analysis of FACL4 on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA12612, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: HEPG2 cell Lane 2: U87-MG cell Lane 3: MC-38 cell Lane 4: U251 cell Lane 5: HELA cell Lane 6: SW620 cell Lane 7: NIH3T3 cell Lane 8: Jurkat cell Predicted molecular weight:79 kDa Observed molecular weight:79 kDa

	Fig : Immunohistochemical analysis of paraffin-embedded Mouse-spleen tissue with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig: Fluorescence immunohistochemical analysis of Rat-urinary bladder tissue (Formalin / PFA-fixed paraffin-embedded sections). with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig : Western blot analysis of FACL4 on different lysates. Proteins were transferred to a NC membrane and blocked with 5% NF-Milk in TBST for 1 hour at room temperature. The primary antibody (AWA12612, 1/1000) was used in TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (AWS0002) at 1:5,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: PC12 cell Lane 2: L6 cell Predicted molecular weight:79 kDa Observed molecular weight:79 kDa

	Fig: Fluorescence immunohistochemical analysis of Mouse-testicle tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig：Flow cytometric analysis of HEK-293T cells labeling FACL4. Overlay histogram showing HEK-293T cells stained with FACL4 (green line). The cell were fixed in 4% paraformaldehyde for 30 minutes at 37 ℃, permeabilized with 0.02% Triton X-100 in PBS for 30 minutes,and then stained with the primary antibody(AWA12612, 1:50 ) for 30 min at 4°C. The secondary antibody used was an Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody (AWS0005b) at 1/2500 dilution for 30 min at 4ºC. Unlabelled sample was used as a control (cells without incubation with primary antibody; red).

	Fig: Fluorescence immunohistochemical analysis of Mouse-cerebellum tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig: Fluorescence immunohistochemical analysis of Rat-uterus tissue (Formalin / PFA-fixed paraffin-embedded sections). with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig: Fluorescence immunohistochemical analysis of Mouse-lung tissue (Formalin/PFA-fixed paraffin-embedded sections). with Rabbit anti-FACL4 antibody (AWA12612) at 1/200 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/200 dilution for 2 hour at 37℃or overnignt at 4℃. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig: Fluorescence immunohistochemical analysis of Rat-brain tissue (Formalin / PFA-fixed paraffin-embedded sections). with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The immunostaining was performed with the TSA Immuno-staining Kit (ABIOWELL, AWI0689). The section was pre-treated using heat mediated antigen retrieval with EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system followed by a separate fluorescent tyramide signal amplification system (red). DAPI (blue, AWC0291) was used as a nuclear counter stain. Image acquisition was performed with Slide Scanner.

	Fig : Immunohistochemical analysis of paraffin-embedded Rat-lung tissue with Rabbit anti-FACL4 antibody (AWA12612) at 1/100 dilution. The section was pre-treated using heat mediated antigen retrieval with Sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 3% H₂O₂ for 15 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (AWA12612) at 1/100 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system(ABIOWELL, AWI0629). DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

引用文献 (3)

BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE IF:4.2

IGF2BP1 exacerbates neuroinflammation and cerebral ischemia/reperfusion injury by regulating neuronal ferroptosis and microglial polarization

Background Cerebral ischemia/reperfusion (I/R) injury induces neuronal ferroptosis and microglial phenotypic shifts, driving post-ischemic neurological deficits. This study examines the regulatory role of the N6-methyladenosine (m6A) reader insulin-like growth factor 2 mRNA-binding protein 1 (IGF2BP1) in coordinating these pathological processes through Keap1/Nrf2 signaling. Methods Cerebral I/R injury was modeled in C57BL/6 mice via middle cerebral artery occlusion (MCAO) and in hippocampal neurons and microglia through oxygen-glucose deprivation/reperfusion (OGD/R). Pro-inflammatory microglial polarization was induced by LPS/IFN-γ stimulation. IGF2BP1's functional impacts were assessed through knockdown and overexpression approaches, with mechanistic evaluations focusing on ferroptosis biomarkers, microglial polarization states, and Keap1/Nrf2 pathway activity. A microglia-neuron co-culture system elucidated cellular crosstalk mechanisms. Results MCAO-operated mice demonstrated upregulated IGF2BP1 expression accompanied by neuronal apoptosis and microglial M1 polarization. IGF2BP1 silencing significantly attenuated OGD/R-induced neuronal ferroptosis, evidenced by reduced iron overload (Fe 2+ ), lipid peroxidation (MDA), and reactive oxygen species (ROS) alongside restored glutathione (GSH) levels, while concurrently enhancing GPX4 activity through Keap1/Nrf2 pathway regulation. This intervention further shifted microglial polarization toward the M2 phenotype, effectively mitigating neuroinflammatory responses. Importantly, the neuroprotective effects of IGF2BP1 knockdown were abolished upon Keap1 overexpression. Co-culture experiments revealed that IGF2BP1-depleted microglia suppressed neuronal ferroptosis via phenotypic reprogramming. In vivo validation confirmed that IGF2BP1 knockdown ameliorated neurological deficits and reduced ferroptosis markers in MCAO-challenged mice. Conclusion IGF2BP1 serves as a critical regulator of cerebral I/R injury by exacerbating neuronal ferroptosis and sustaining detrimental microglial activation. These findings nominate IGF2BP1 inhibition as a promising strategy for ischemic stroke intervention.

pubTime 2025-04-26

Application

Specie

Mouse

Dilution

1:1000

JOURNAL OF NEUROCHEMISTRY IF:4

Exosome-Derived CDC42 From Hypoxia-Pretreated Neural Stem Cells Inhibits ACSL4-Related Ferroptosis to Alleviate Vascular Injury in Parkinson's Disease Mice Models

Parkinson's disease (PD) is a neurodegenerative disorder that gets exacerbated by vascular injury. Neural stem cell-derived exosomes (NSC-Exos) display effective neuroprotective properties in PD models. Cell division control protein 42 (CDC42) is connected to angiogenesis, but its effects in PD remain undefined. This research aims to reveal the role of CDC42 in PD. First, we applied 1-methyl-4-phenylpyridinium (MPP + ) to induce the human cerebral microvascular endothelial cells (HCMECs) model and evaluated cell viability and ferroptosis. Then, we characterized NSC-Exos. Next, to appraise the effect of hypoxia-pretreated NSC-Exos (H-NSC-Exos) on the MPP + -induced cells model, we examined angiogenesis and ferroptosis in HCMECs. Moreover, we constructed the PD mice model using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and tested the behavioral experiments and vascular injury of mice. Furthermore, we examined cellular ferroptosis and angiogenesis after knockdown of CDC42. Additionally, we investigated the interaction of CDC42 with Acyl-CoA synthetase long-chain family member 4 (ACSL4) and detected cellular ferroptosis and angiogenesis after overexpression of ACSL4. We found that H-NSC-Exos reversed the MPP + -induced decrease in HCMECs viability and migration, lowered lipid-reactive oxygen species (lipid-ROS) levels, suppressed ferroptosis, and facilitated angiogenesis. Moreover, H-NSC-Exos attenuated MPTP-induced PD development, vascular injury, and ferroptosis in mice. H-NSC-Exos with the knockdown of CDC42 reduced cell viability and angiogenesis and raised ferroptosis and lipid-ROS levels, which were reversed by ferrostatin-1 and liproxstatin-1. CDC42 interacted with ACSL4. Furthermore, overexpression of ACSL4 aggravated the above effects of H-NSC-Exos in which CDC42 was knocked down. Our study reveals that H-NSC-Exos-derived CDC42 inhibited ACSL4-related ferroptosis to alleviate vascular injury in PD mice models. CDC42 may serve as a potent therapeutic target for PD treatment.

pubTime 2025-03-04

Application

WB,CoIP

Specie

Human

Dilution

1:1000

CELLULAR SIGNALLING IF:3.7

Set7 regulates ferroptosis in pulmonary arterial hypertension endothelial cells through the DPP4/NOX4 axis mediated by H3K4me1 modification

Objective Dipeptidyl peptidase-4 (DPP4)-targeted therapy is widely employed in the therapy of pulmonary diseases, but the role of its H3K4me1 modification in pulmonary arterial hypertension (PAH) disease remains unknown. This study aims to investigate the function of histone methyltransferase SET domain containing 7 (Set7)-mediated monomethylation of histone 3 lysine 4 (H3K4me1) modification of DPP4 in PAH, with the goal of providing new insights for the broader application of DPP4-targeted therapies. Methods The PAH mouse model was constructed and intervened with overexpression (oe) or knockdown (sh) of DPP4, sh-Set7, oe-NADPH oxidase 4 (NOX4) or sh-Set7 + erastin. Human pulmonary arterial endothelial cells were induced by hypoxia and treated with sh-DPP4, erastin, oe-DPP4, sh-Set7, oe-NOX4, oe-Set7 or oe-Set7 + ferrostatin-1. The enrichment of H3K4me1 level in the DPP4 promoter region was analyzed by ChIP and dual-luciferase assay. Pulmonary vascular remodeling, fibrosis, and endothelial injury were observed by echocardiography, HE, MASSON, and α-SMA staining. Ferroptosis markers and protein expression were measured using biochemical assay kits, RT-qPCR, WB, immunofluorescence, and transmission electron microscopy. Results Silencing DPP4 alleviates pulmonary vascular remodeling, fibrosis, and endothelial injury in PAH mice, reduces cardiac fibrosis and pulmonary inflammation, while improving mitochondrial damage in the lungs and downregulating the level of ferroptosis-related proteins. ChIP assays confirmed increased enrichment of H3K4me1 in the DPP4 promoter region in both hypoxia-induced endothelial cells and lung tissues of PAH mice. Overexpression of Set7 resulted in elevated H3K4me1 enrichment in the DPP4 promoter region and increased NOX4 protein expression. Ferrostatin-1 inhibited the promotion of oe-Set7 in hypoxia-induced endothelial cell injury. Silencing Set7 mitigated hypoxia-induced endothelial cell injury, ferroptosis, and inflammatory responses by downregulating DPP4/NOX4. Erastin reversed the treatment effect of sh-Set7 in PAH mice. Furthermore, Set7 knockdown ameliorated PAH in mice by suppressing DPP4/NOX4-mediated ferroptosis. Conclusion The H3K4me1 modification of DPP4 is upregulated in PAH, a process regulated by Set7. Silencing Set7 alleviates PAH by suppressing ferroptosis through the DPP4/NOX4 signaling pathway, offering a novel gene therapy approach for this disease.

pubTime 2026-01-05

Application

Specie

Mouse,Human

Dilution

1:1000