Ab200353

Manufactured by Abcam

Ab200353 is an antibody that can be used for laboratory applications. It is a purified rabbit polyclonal antibody.

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Lab products found in correlation

4 protocols using ab200353

Western Blot Analysis of Fatty Acid Transporters

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Liver (40 mg) or subcutaneous fat (100 mg) was homogenized using a Bullet Blender. Lysates from cultured MΦ (30 µg), liver homogenates (30 µg), or visceral fat homogenates (80 µg) were separated by gel electrophoresis and transferred onto nitrocellulose or PVDF membranes. Membranes were incubated overnight with an antibody against SLC27A4/FATP4 (ab200353, Abcam) or CEBPα (clone EP709Y, Cat. No. 1704-1, Epitomics). Primary antibodies purchased from Cell Signaling (Frankfurt, Germany) were antibodies against MCP-1 (Cat. No. 2029), PPARγ (Cat. No. 2435), p-FoxO1 (Ser256; Cat. No. 9461), ACC (Cat. No. 3662), FASN (C20G5, Cat. No. 3180), and GAPDH (14C10, Cat. No. 2118). After incubation with a secondary HRP-linked antibody, proteins were visualized by using Luminata Forte Western HRP Substrate (Millipore, Darmstadt, Germany). Quantification of proteins was carried out using ImageJ after normalization to GAPDH.

Göcebe D., Jansakun C., Zhang Y., Staffer S., Tuma-Kellner S., Altamura S., Muckenthaler M.U., Merle U., Herrmann T, & Chamulitrat W. (2023). Myeloid-specific fatty acid transport protein 4 deficiency induces a sex-dimorphic susceptibility for nonalcoholic steatohepatitis in mice fed a high-fat, high-cholesterol diet. American Journal of Physiology - Gastrointestinal and Liver Physiology, 324(5), G389-G403.

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Immunofluorescence Analysis of FATP4 in ECFCs

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ECFCs overexpressing FATP4 were fixed in 3.7% paraformaldehyde for 10 min. After fixation, the cells were permeabilized in 0.3% Triton X-100 for 5 min and blocked with 3% BSA for 30 min. For Figures 4E–F, the cells were then incubated with primary antibodies overnight at 4°C: anti-COX4-I1 (R&D, AF5814), anti-KDEL (Abcam, ab12223), and anti-FATP4 (Abcam, ab200353). For Figure S4H, cells were incubated with anti-FATP4 and anti-CDH5 (Thermofisher Scientific, 12-1449-82). After washing with PBS, the cells were incubated with secondary antibodies for two hours at room temperature. After washing with PBS, cells were mounted with VectaShield mounting medium (VectorLabs). Imaging was done using the Zeiss LSM 880 with Fast Airyscan module (laser excitation 405 nm for KDEL, 488nm for COX4-I1 (and CDH5), and 560nm for FATP4). Image processing and the intensity values of the line profile analysis were done by using Zen 3.0 blue software.

Ibrahim A., Yucel N., Kim B, & Arany Z. (2020). Local Mitochondrial ATP Production Regulates Endothelial Fatty Acid Uptake and Transport.. Cell metabolism, 32(2), 309-319.e7.

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Lipid Metabolism Protein Analysis

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SOCS5 (1:500, sc-100858, Santa Cruz); HA-Tag (1:1000, #3724, CST); hnRNP G (1:1000, ab190352, Abcam); FASN (1:1000, #3180, CST); Myc-Tag (1:1000, #2276, CST); ACC (1:1000, #3676, CST); SCD1 (1:1000, ab236868, Abcam); ACLY (1:5000, ab40793, Abcam); SREBP1 (1:1000, 14088-1-AP, Proteintech); IgG (1:1000, #2729, CST); HRP anti-mouse IgG (1:10,000, abs20001, Absin); HRP anti-rabbit IgG (1:10,000, abs20002, Absin); CD36 (1:1000, ab252922, Abcam); FATP4 (1:1000, ab200353, Abcam).

Wang Y., Zhao Z., Guo T., Wu T., Zhang M., Luo D., Dou K., Yang Y., Jin C., Zhang B., Zhang B, & Han B. (2024). SOCS5-RBMX stimulates SREBP1-mediated lipogenesis to promote metastasis in steatotic HCC with HBV-related cirrhosis. NPJ Precision Oncology, 8, 58.

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Western Blot Analysis of Adipose Tissue

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The homogenates of scWAT and BAT were dissolved in RIPA lysate and protease inhibition for protein extraction. Sample protein concentrations were measured using the bicinchoninic acid (BCA) method (Beyotime). Total protein (10μg/Lane) was separated on a 12% acrylamide/acrylamide gel using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene difluoride (PVDF) membranes. PVDF membranes containing protein were incubated with specific anti-alpha tubulin antibody (1:5000, ab18251, Abcam), anti-UCP1 antibody (1:1000, ab10983, Abcam), anti-Prohibitin antibody (1:10000, ab75766, Abcam), anti-PPAR α antibody (1:2000, ab8934, Abcam), anti-CD36 antibody (1:5000, ab133625, Abcam), anti-slc27a4/FATP4 antibody (1:1000, ab200353, Abcam), respectively. Then membranes were incubated with HRP-conjugated Affinipure Goat Anti-Mouse IgG (H+L) (1:5000, 20000175, proteintech) or HRP-conjugated Affinipure Goat Anti-Rabbit IgG (H+L) (1:5000, 20000174, proteintech). Protein bands were visualized using the ECL kit (EMD millipore). Image was analyzed using Image-Pro-Plus 6.0.

Cheng L., Zhang S., Shang F., Ning Y., Huang Z., He R., Sun J, & Dong S. (2021). Emodin Improves Glucose and Lipid Metabolism Disorders in Obese Mice via Activating Brown Adipose Tissue and Inducing Browning of White Adipose Tissue. Frontiers in Endocrinology, 12, 618037.

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