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Ab54430

Manufactured by Abcam
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Ab54430 is a laboratory equipment product offered by Abcam. It serves as a core component for various research applications, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

Alexa fluor 568, by Thermo Fisher Scientific (2 mentions) Mca1957, by Bio-Rad (2 mentions) Mca497ga, by Bio-Rad (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Ba 2000, by Vector Laboratories (2 mentions) Af1109, by R&D Systems (2 mentions) Sa 5008, by Vector Laboratories (2 mentions) Ba 9500, by Vector Laboratories (2 mentions) Universal magnetic co ip kit, by Active Motif (1 mentions) Lipofectamine 3000 reagent, by Thermo Fisher Scientific (1 mentions) Ab32049, by Abcam (1 mentions) Antibody against flag f1804, by Merck Group (1 mentions) Overexpression lentivirus, by Obio Technology (1 mentions) Ab32124, by Abcam (1 mentions)

3 protocols using ab54430

1

UNC5B Overexpression Lentivirus Protocol

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UNC5B(399‐945aa)‐3Flag overexpression lentivirus and UNC5B(412‐945aa)‐3Flag overexpression lentivirus were synthesized by Obio Technology Co. Ltd. Tables 1 and 2 indicated the composition reports. Full‐length UNC5B‐Flag overexpression lentivirus was purchased from GenePharm Co. Ltd. The Lipofectamine 3000 Reagent (Life Technologies Co. Ltd.) was used for these plasmids and related transfection.
Antibodies against UNC5B (ab104871, ab54430), caspase‐3 (ab179517), mutant P53 (ab32049) and BCL‐2 (ab32124) were purchased from Abcam. Antibodies against cleaved PARP (D64E10), S6 Ribosomal Protein (5G10), UNC5B (D9M7Z) and beta‐actin (3700S) were obtained from Cell Signaling Technology (CST). Antibodies against Fibronectin, beta‐catenin, vimentin, N‐CA and E‐CA were also obtained from the Epithelial‐Mesenchymal Transition (EMT) Antibody Sampler Kit (9782T, CST). Antibody against FLAG (F1804) was obtained from Sigma‐Aldrich Co. Ltd. Antibody against P53 (21891‐1) was purchased from Proteintech Co. Ltd. UNC5B (ab54430, Abcam) was used for immunofluorescence and flow cytometry. UNC5B (D9M7Z) was used as a mixture reagent in Co‐IP. Universal Magnetic Co‐IP Kit (54002, Active Motif) was purchased from Active Motif Co. Ltd. Fast silver staining kit (P0017S, Beyotime) was purchased from Beyotime Co. Ltd.
Huang Y., Zhang Z., Miao M, & Kong C. (2020). The intracellular domain of UNC5B facilities proliferation and metastasis of bladder cancer cells. Journal of Cellular and Molecular Medicine, 25(4), 2121-2135.
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2

Immunofluorescence Analysis of Adipose Tissue

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White adipose tissue was excised, fixed in formalin overnight, embedded in paraffin and sectioned. The immunofluorescence analysis of netrin-1 (1:200, AF1109; R&D systems), caveolin-1 (1:500, 610059; BD bioscience), Unc5b (1:200, ab54430; Abcam), F4/80 (1:500, MCA497GA; Abd serotec), CD68 (1:500, MCA1957; Abd serotec) was conducted after deparaffinization as described previously61 (link). Secondary antibodies were then applied (Alexa Fluor-568, A11011; Alexa Fluor-488, A11006; 1:500 Invitrogen). Netrin-1 and Unc5b staining was amplified using the biotin conjugated antibodies (1:100, BA-2000 and BA-9500; Vector Laboratories) followed by streptavidin conjugated AMCA (1:200, Sa-5008; Vector Laboratories) staining. Sections were mounted and visualized using a Nikon Eclipse microscope.
Ramkhelawon B., Hennessy E.J., Ménager M., Ray T.D., Sheedy F.J., Hutchison S., Wanschel A., Oldebeken S., Geoffrion M., Spiro W., Miller G., McPherson R., Rayner K.J, & Moore K.J. (2014). Netrin-1 promotes adipose tissue macrophage accumulation and insulin resistance in obesity. Nature medicine, 20(4), 377-384.
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3

Immunofluorescence Analysis of Adipose Tissue

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White adipose tissue was excised, fixed in formalin overnight, embedded in paraffin and sectioned. The immunofluorescence analysis of netrin-1 (1:200, AF1109; R&D systems), caveolin-1 (1:500, 610059; BD bioscience), Unc5b (1:200, ab54430; Abcam), F4/80 (1:500, MCA497GA; Abd serotec), CD68 (1:500, MCA1957; Abd serotec) was conducted after deparaffinization as described previously61 (link). Secondary antibodies were then applied (Alexa Fluor-568, A11011; Alexa Fluor-488, A11006; 1:500 Invitrogen). Netrin-1 and Unc5b staining was amplified using the biotin conjugated antibodies (1:100, BA-2000 and BA-9500; Vector Laboratories) followed by streptavidin conjugated AMCA (1:200, Sa-5008; Vector Laboratories) staining. Sections were mounted and visualized using a Nikon Eclipse microscope.
Ramkhelawon B., Hennessy E.J., Ménager M., Ray T.D., Sheedy F.J., Hutchison S., Wanschel A., Oldebeken S., Geoffrion M., Spiro W., Miller G., McPherson R., Rayner K.J, & Moore K.J. (2014). Netrin-1 promotes adipose tissue macrophage accumulation and insulin resistance in obesity. Nature medicine, 20(4), 377-384.
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