rabbit anti aplp2 antibody  (Boster Bio)

Journal: medRxiv

Article Title: The congenital multiple organ malformation syndrome, Ritscher-Schinzel syndrome is an endosomal recyclinopathy

doi: 10.1101/2024.08.17.24311658

Figure Lengend Snippet: SNX17-Retriever/CCC/WASH pathway is essential for recycling of Reelin signaling receptors, APP family, and SLITRK family proteins. (A) Representative blots of co-expression of mCherry-tagged SNX17 with GFP-tagged cytoplasmic tail of LRP8 and VLDLR from three independent experiments. The interactions between mCherry-SNX17 and wild-type GFP-tagged cytoplasmic tails of Human-LRP8 and Human-VLDLR were significantly decreased when NPxY was substituted with NPxA. (B) Representative blots of analysis from three independent experiments using DIV17 rat cortical neurons transduced with either a scramble-control, SNX17, or VPS35L shRNA. Bar graphs show relative protein abundance of LRP8 and VLDLR in cell lysate or cell surface. (C) DIV17 rat cortical neurons transduced with shRNA were incubated for 30 min with or without AP-Reelin, followed by cell lysis and western blot analysis. Bar graph shows quantification of band intensities of AP relative to cells transduced with sh-SCR control from three independent experiments. (D) DIV17 rat cortical neurons transduced with shRNA were incubated for 7 min with or without Reelin. Phosphorylation level of Dab1 was then measured using immunoprecipitation and western blot analysis. Representative blots and quantification from three independent analyses are shown. (E) Volcano plots of transmembrane proteins with decreased (blue circles) or increased (red circles) cell surface abundance in sh-VPS35L suppression comparedto sh-SCR in DIV17 rat cortical neuron from three independent TMT-based proteomic experiments. Two independent sh-RNAs for VPS35L were used to avoid off-target effects. (F) Enrichment analysis of significantly downregulated proteins (LogFC < −0.32, p < 0.05) in the sh-VPS35L compared to sh-SCR by Metascape. (G) Representative blots of mCherry-nanotrap for mCherry-SNX17 under co-overexpression of chimeric constructs of Human-SLITRK family proteins from three independent experiments. All proteins except for SLITRK4 have an NPxY motif, which was mutated to NPxA in the mutant. (H) Representative blots of mCherry-nanotrap for mCherry-SNX17 under co-overexpression of chimeric constructs of Human-APP family proteins from three independent experiments. APP, APLP1, and APLP2 have NPxY motifs, and the NPxY motif was mutated to NPxA in the mutant. (I) Representative blots for APP and APLP2 in rat cortical neurons. Cell surface protein fraction was obtained, and N-Cadherin were used for loading control. Bar graphs show relative values of cells with sh-SNX17 or sh-VPS35L suppression compared to sh-SCR control cells (n=3). In all graphs, error bars represent mean ± SD. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001.

Article Snippet: The following antibodies were used in this study (WB: western blot, IF: immunofluorescence): rabbit anti-SNX17 (Proteintech, 10275-1-AP, WB), mouse anti-GFP (Roche, 11814460001, WB), rabbit anti-GFP (GeneTex, GTX30738, WB), mouse anti-mCherry (antibodies.com, A85305, WB), rabbit anti-mCherry (antibodies.com, A85306, WB), rabbit anti-CCDC22, (Proteintech, 16636-1-AP, WB), mouse anti-CCDC93, (Origene, CF800568, WB), rabbit anti COMMD4 and rabbit anti COMMD9 (kind gift from Prof. Ezra Burstein, WB) rabbit anti-C16orf62 (Abcam, ab97889, WB), rabbit anti-C16orf62 (Pierce, PA5-28553, IF), rabbit anti-DSCR3 (Merck Millipore, ABN87, WB), rabbit anti-integrin-β1 (Abcam, ab52971, WB), goat anti-VPS35 (antibodies.com, A83699, IF), mouse anti-VPS29 (Santa Cruz, sc-398874, WB), rabbit anti-KIAA1033 (Proteintech, 51101-1-AP, WB), mouse anti-Strumpellin (Santa Cruz, sc-377146, WB), mouse anti-β actin (Sigma, A1978, WB), rabbit anti-LRP1 (Abcam, ab92544, WB), rabbit anti-LRP2 (Proteintech, 19700-1-AP, WB), rabbit anti-LRP2 (prepared as described in , IHC), rabbit anti-LRP4 (SIGMA, HPA012300, WB), rabbit anti-LRP8 (Abcam, ab108208, WB), mouse anti-VLDLR (Santa Cruz, sc-18824, WB), rabbit anti-APP (Abcam, ab32136, WB), rabbit anti-APLP2 (Proteintech, 15041-1-AP, WB), mouse anti-AP (Thermo Fisher, MA1-20245, WB), rabbit anti-Dab1 (kind gift from Dr. M Hattori, IP/WB) , mouse anti-Phosphotyrosine (Merck, 05-321, WB), rabbit anti-GLUT1 (Abcam, ab115730, IF/WB), mouse anti-N-cadherin (Cell signalling technology, 14215S, WB), anti-PSD95 (Merck, MAB1596, WB), mouse anti-FLAG (SIGMA, F1804, WB), rabbit anti-FGFR2, (Proteintech, 13042-1-AP, WB), rabbit anti-ERK1/2 (Cell Signaling Technology, 9102, WB), rabbit anti-phospho-ERK1/2 (Cell Signaling Technology, 9101, WB), anti Ctip2 (Abcam, ab18465, IHC), anti Tbr1 (Abcam, ab275960, IHC), anti Neun (Cell signalling technology, 12943, IHC).

Techniques: Expressing, Transduction, Control, shRNA, Incubation, Lysis, Western Blot, Immunoprecipitation, Over Expression, Construct, Mutagenesis

Journal: Molecular Neurodegeneration

Article Title: APP-BP1 inhibits Aβ42 levels by interacting with Presenilin-1

doi: 10.1186/1750-1326-2-3

Figure Lengend Snippet: APP-BP1 expression increased APP processing in primary neurons . A . Expression of myc-tagged APP-BP1. Equal amount of total proteins from neurons infected with APP-BP1 virus or p1005+ virus was analyzed by western blotting using the mouse ant-myc antibody, 9E10. B . APP-BP1 overexpression downregulated rat endogenous APP. APP expression from 15 μg of proteins was analyzed by immunoblots using the anti-APP antibody 369. The amount of γ-tubulin from the same blot was reprobed with a mouse anti-γ-tubulin antibody. Representative blots from the same experiments are shown. C . Quantitative western blot analyses revealed that APP-BP1 significantly downregulated the endogenous APP but not APLPs in neurons overexpressing myc APP-BP1. mycAPP-BP1 expression significantly decreased APP levels compared to the sample infected with the vector (p1005+) (n = 4, p < 0.02, one-tail t -Test). APLP1 levels stayed the same, and APLP2 did not showed a significant change by t -Tests (n = 3, p = 0.1, one tail t -Test). D . RIPA buffer soluble protein extracts from primary neurons co-expressing APP-BP1 and APP 695 or p1005+ and APP 695 were precipitated with 6E10 followed by western blot analyses using 4G8. Mean levels of Aβ is presented in the graph (n = 4, p < 0.04). A representative of 4 independent blots was shown as an insert.

Article Snippet: The following antibodies were used: rabbit anti-PS1 (AB5308) and 22C11 (Chemicon), BP339 [ ], mouse anti-APP-BP1 (BD Transduction Lab), 6E10 and 4G8 (Signet), rabbit anti-cyclin B1 (Santa Cruz), 9E10 (ATCC), rabbit anti-APLP1 and rabbit anti-APLP2 (both from Calbiochem), rabbit anti-nicastrin, rabbit anti-β-catenin and mouse anti-γ-tubulin (all three from Sigma).

Techniques: Expressing, Infection, Western Blot, Over Expression, Plasmid Preparation

Journal: Neuron

Article Title: APP Family Regulates Neuronal Excitability and Synaptic Plasticity but Not Neuronal Survival

doi: 10.1016/j.neuron.2020.08.011

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit anti-APLP2-C-term [752–763] , Millipore , AB_211446.

Techniques: Plasmid Preparation, Virus, Recombinant, Protease Inhibitor, Blocking Assay, Electron Microscopy, Membrane, In Situ, Transgenic Assay, Software