Rabbit anti fip200

Manufactured by Proteintech

Rabbit anti-FIP200 is a primary antibody that recognizes the FIP200 protein. FIP200 is a protein involved in autophagy regulation. This antibody can be used for various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry, to study the expression and localization of FIP200 in biological samples.

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Anti-FIP200 FIP200 Anti-TM

3 protocols using rabbit anti fip200

Immunofluorescence Staining of Autophagy-Related Proteins

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Protocols have been extensively described recently (Lucocq et al., 2001 (link), Karanasios and Ktistakis, 2015 (link)). In brief, cells on coverslips were fixed in 3.7% Formaldehyde, permeabilized in 0.1% NP40 and stained in a blocking solution containing fish gelation and 0.05% NP40. Antibodies used and their final dilution are as follows: rabbit anti-OPTN (Cayman Chemicals), 1:100; mouse anti-WIPI2 (Bio-Rad), 1:200; rabbit anti-phospho(S172)TBK1 (Cell Signalling), 1:50; mouse anti-Ubiquitin FK2 (Enzo), 1:100; rabbit anti-FIP200 (ProteinTech), 1:100; rabbit anti-ATG9 (Cell Signalling), 1:100; mouse anti-TOMM20 (Abcam), 1:100; rabbit anti-ATG13 (Sigma), 1:100; rabbit anti-LC3 (Sigma), 1:150; rabbit anti-NDP52 (GeneTex), 1:100; rabbit anti-Tax1BP1 (ProteinTech), 1:100; rabbit anti-VAPA (Sigma), 1:200; rabbit anti-VAPB (Sigma), 1:200; mouse anti-EEA1 (BD Biosciences), 1:70; mouse anti-LAMP2 (Developmental Studies Hybridoma Data Bank), 1:200; mouse anti-TUBULIN (Sigma), 1:300; mouse anti-cytochrome C (Abcam), 1:200; rabbit anti-phospho(S351)p62 (kind gift from Dr Masaaki Komatsu), 1:100; rabbit anti-TRAF2 (Abcam) 1:100.

Zachari M., Gudmundsson S.R., Li Z., Manifava M., Shah R., Smith M., Stronge J., Karanasios E., Piunti C., Kishi-Itakura C., Vihinen H., Jokitalo E., Guan J.L., Buss F., Smith A.M., Walker S.A., Eskelinen E.L, & Ktistakis N.T. (2019). Selective Autophagy of Mitochondria on a Ubiquitin-Endoplasmic-Reticulum Platform. Developmental Cell, 50(5), 627-643.e5.

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Immunofluorescence Staining of Neural Markers

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Primary antibodies used were: mouse anti-BrdU (G3G4; DSHB), mouse anti-GFAP (Thermo Fisher Scientific), rhodamine-conjugated mouse anti-BrdU (Millipore), mouse anti-nestin (Rat-401), mouse anti-NeuN (Millipore), mouse anti–Nqo-1 (Santa Cruz Biotechnology), mouse anti-p62 (Abcam), mouse anti ubiquitin (Santa Cruz Biotechnology), mouse antivinculin (Sigma-Aldrich), rabbit anti-Atg5 (Novus Biologicals), rabbit anti-Atg16L1 (Abgent), rabbit anti-GFAP (Dako), rabbit anti-Ki67 (Spring Bioscience), rabbit anti-LC3 (Cell Signaling), rabbit anti-NBR1 (Cell Signaling), rabbit anti-Nrf2 (Abcam), rabbit anti-p62 (Enzo), rabbit anti-Sox2 (Millipore), rabbit anti-TAX1BP1 (Abgent), rabbit anti-Fip200 (ProteinTech), rabbit anti-olig2 (Millipore), rat anti-Ki67 (BioLegend), and guinea pig antidoublecortin (anti-DCX; EMD Millipore). Secondary antibodies were goat anti–rabbit IgG-FITC, goat anti–rabbit IgG–Texas red, goat anti–mouse IgG-FITC, goat anti–mouse IgG–Texas red, goat anti–mouse IgG-HRP, and goat anti–rabbit IgG-HRP (Jackson Immunology).
DHE and EUK-8 were purchased from Sigma-Aldrich. EUK-134 was purchased from Cayman Chemical. DCFDA, MitoTracker Red, and MitoTracker Green were purchased from Invitrogen.

Wang C., Chen S., Yeo S., Karsli-Uzunbas G., White E., Mizushima N., Virgin H.W, & Guan J.L. (2016). Elevated p62/SQSTM1 determines the fate of autophagy-deficient neural stem cells by increasing superoxide. The Journal of Cell Biology, 212(5), 545-560.

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Autophagy Protein Immunodetection Assays

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The following antibodies were used for immunostaining or immunoblotting assays: mouse anti-LC3 (MBL, M152-3), rabbit anti-LC3 (2775S; Cell Signaling Technology), rabbit anti-p62 (MBL, PM045), rabbit anti-FIP200 (SAB4200135; Sigma-Aldrich), rabbit anti-FIP200 (17250-1-AP; Proteintech), rabbit anti-ULK1 (8054; Cell Signaling Technology), rabbit anti-Atg13 (13468S; Cell Signaling Technology), rabbit anti-ATL2 (16688-1-AP; Proteintech), rabbit anti-ATL3 (ab117819; Abcam), mouse anti-WIPI2 (ab105459; Abcam), mouse anti-VAPB (66191-1-IG; Proteintech), rabbit anti-VAPA (15275-1-AP; Proteintech), mouse anti-LAMP1 (553792; BD Biosciences), rabbit anti-PTPIP51 (20641-1-AP; Proteintech), rabbit anti-ATG101 (13492; Cell Signaling Technology), rabbit anti-Myc (2278S; Cell Signaling Technology), mouse anti-Myc (M5546; Sigma-Aldrich), mouse anti-GFP (11814460001; Roche), mouse anti-actin (60008-1-IG; Proteintech), and rabbit anti-HA (H6908; Sigma-Aldrich).
The following reagents were used in this study: wortmannin (PHZ1301; Life Technologies), bafilomycin (B1793; Sigma-Aldrich), rapamycin (R0395; Sigma-Aldrich), Lipofectamine 2000 (12566014; Life Technologies), Lipofectamine RNAi MAX (13778150; Life Technologies), and LysoTracker Deep Red (L12492; Thermo Fisher Scientific).

Liu N., Zhao H., Zhao Y.G., Hu J, & Zhang H. (2021). Atlastin 2/3 regulate ER targeting of the ULK1 complex to initiate autophagy. The Journal of Cell Biology, 220(7), e202012091.

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