Dendra 2 Antikörper

Produktnummer ABIN361314

Produktdetails

Highlights:

• Dendra2 Antibody (ABIN361314) independently validated for Immunocytochemistry by Neurology Lab (INSERM).
• Publications cite use for Immunoblotting as well as Immunostaining.
• Dendra2 belongs to a new class of photoactivatable fluorescent proteins (PAFPs). PAFPs allow to photolable and track fusion proteins in vivo. Dendra2 Antibody (ABIN361314) can be used for control purposes in this setting.

Target: Dendra 2

Reaktivität: Koralle

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Unkonjugiert

Applikation: Western Blotting (WB), Flow Cytometry (FACS), Immunocytochemistry (ICC)

Spezifität: The rabbit polyclonal antibody PAb (836) recognizes Dendra2 tag, a green-to red photoswitchable protein derived from an octocoral (Dendronephthya).

Aufreinigung: Purified by protein-A affinity chromatography

Reinheit: > 95 % SDS

Immunogen: Dendra2 tag

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: DENDRA 2 antibody has been tested for use in Western Blotting, Flow Cytometry and Immunocytochemistry. It is expected that the antibody will be suited for all applications where recognition of the native protein is required. A titration should be performed to determine the optimal concentration

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (ICC)

Validierung #101498 (Immunocytochemistry)

by: Plasticity in cortical networks and epilepsy, Institut du Fer à Moulin, UMR839 INSERM/UPMC

No.: #101498

Datum: 03.11.2017

Antigen: Dendra2

Chargennummer: R-053-AI-03

Validierte Anwendung: Immunocytochemistry

Positivkontrolle: Rat hippocampal cells transfected with a Dendra2-tagged protein (Kv2.1_Dendra)

Negativkontrolle: Non transfected cells in the same preparation

Bewertung:

ABIN361314 specifically labels Dendra+ cells in hippocampal cell cultures transfected with a construct bearing Dendra2.

Validierungsbilder

Hippocampal primary culture co-transfected with Kv2.1_Dendra and HomerDsRed (A, red fluorescence of Homer), stained with anti-Dendra2 ABIN361314 (B, revealed with A647-coupled secondary antibody) and anti-MAP2 antibody (C, revealed with A488-coupled secondary antibody). Merged images in (D), showing the labeling of anti-Dendra2 antibody (blue) that reveals the transfected cell identified by the fluorescence of HomerDsRed (red) but not the non-transfected cells (visible by the immunolabeling of MAP2, green). Scale bar: 10µm, 63x magnification.

Protokoll

Primärantikörper: ABIN361314

Sekundärantikörper: donkey anti-rabbit A647 conjugated antibody (Jackson Immunoresearch, 711-605-152, lot 123104)

Full Protocol:

• Grow hippocampal cells in Neurobasal medium 1x (Gibco, ref. 21103-049, lot 1880280) supplemented with B27 1x (Gibco, ref. 17-504-044, lot 1733317), 2mM L-glutamine (Gibco, ref. 25030-024, lot 1881896) and antibiotics (200U/ml penicillin, 200µg/ml streptomycin, Gibco, ref. 15140-122), at 37°C and 5% CO₂ at a density of 120×10³ cells/ml on 18-mm diameter glass coverslips (Assistent) pre-coated with 50µg/ml poly-D,L-ornithine (Sigma-Aldrich, P3655, lot SLBL9424V).
• Transfect cells with 0.5µg of Kv2.1_Dendra and 0.5µg of Homer_DsRed constructs (in-house made) using Transfectin Lipid transfection reagent (BioRad, ref 170-3351, lot 4206491) following the manufacturer´s instructions.
• Fix hippocampal cell cultures on coverslips with 4% buffered formalin for 15min at RT.
• Wash slides 2x in PBS (prepared from DPBS 10x, Gibco, 14200-067, lot 1754645).
• Permeabilize cells with 0.25% Triton X-100 (Roth, 3051.3) in PBS for 4min at RT.
• Wash slides 2x in PBS.
• Block non-specific binding with PBS-Goat serum 10% (Goat serum: Gibco, 16210-064, lot 1158993) for 30min at RT.
• Incubate slides with primary rabbit anti-Dendra2 antibody (antibodies-online, ABIN361314, lot R-053-AI-03) diluted 1:100 and mouse anti-MAP2 antibody (Chemicon, MAB 378, lot LV1414361) diluted 1:200 in PBS-Goat serum 10% for 1h at RT.
• Wash slides 2x 5min in PBS-Goat serum 5%.
• Incubate with secondary donkey anti-rabbit A647 conjugated antibody (Jackson Immunoresearch, 711-605-152, lot 123104) and donkey anti-mouse A488 conjugated antibody (Jackson Immunoresearch, 715-545-150, lot 123589) diluted 1:250 in PBS-Goat serum 10% for 45min at RT.
• Wash slides 1x 5min in PBS.
• Mount coverslips with 10-20µl of Mowiol antifade reagent (Calbiochem, 475904, lot D00136768).
• Image acquisition on a Nikon Eclipse Ti microscope, 63x magnification.

Anmerkungen:

Staining pattern of Kv2.1_Dendra observed with ABIN361314 reflects the staining of the native protein Kv2.1 (obtained with its specific antibody).

Handhabung

Konzentration: 1mg/ml

Buffer: Phosphate buffered saline (PBS) solution with 15 mM sodium azide

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handhabung: Do not freeze.

Lagerung: 4 °C

Informationen zur Lagerung: Store at 2-8°C.

Referenzen

Song, Ma, Liu, Huang, Li, Li, Luo, Ni, He: "The MRN complex maintains the biliary-derived hepatocytes in liver regeneration through ATR-Chk1 pathway." in: NPJ Regenerative medicine, Vol. 8, Issue 1, pp. 20, (2023) (PubMed).

Morgner, Bornes, Hahn, López-Iglesias, Kroese, Pritchard, Vennin, Peters, Huijbers, van Rheenen: "A Lamb1Dendra2 mouse model identifies basement-membrane-producing origins and dynamics in PyMT breast tumors." in: Developmental cell, Vol. 58, Issue 7, pp. 535-549.e5, (2023) (PubMed).

Zhang, Zhou, Li, Mo, Ma, Ni, Yang, He, Luo: "Tel2 regulates redifferentiation of bipotential progenitor cells via Hhex during zebrafish liver regeneration." in: Cell reports, Vol. 39, Issue 1, pp. 110596, (2022) (PubMed).

Wang, Fan, Khawaja, Liu, Zhan, Kodama, Chin, Li, Le, Zhou, Condello, Grinberg, Seeley, Miller, Mok, Gestwicki, Cuervo, Luo, Gan: "Microglial NF-κB drives tau spreading and toxicity in a mouse model of tauopathy." in: Nature communications, Vol. 13, Issue 1, pp. 1969, (2022) (PubMed).

Singh, Ye, Kim: "Dual leucine zipper kinase regulates Dscam expression through a non-canonical function of the cytoplasmic poly(A)-binding protein." in: The Journal of neuroscience : the official journal of the Society for Neuroscience, (2022) (PubMed).

Chen, He, Luo: "Brain vascular damage-induced lymphatic ingrowth is directed by Cxcl12b/Cxcr4a." in: Development (Cambridge, England), Vol. 149, Issue 13, (2022) (PubMed).

Singh, Kim: "Generation and validation of pX-UASTattB for dose-dependent misexpression studies in Drosophila." in: microPublication biology, Vol. 2022, (2022) (PubMed).

Teh, Singh, Ren, Huang, Ariyarathne, Salamon, Wang, Kotake, Fujita: "Surface-localized glycoproteins act through class C ARFs to fine-tune gametophore initiation in Physcomitrium patens." in: Development (Cambridge, England), Vol. 149, Issue 24, (2022) (PubMed).

Bellec, Pinot, Gicquel, Le Borgne: "The Clathrin adaptor AP-1 and Stratum act in parallel pathways to control Notch activation in Drosophila sensory organ precursors cells." in: Development (Cambridge, England), Vol. 148, Issue 1, (2021) (PubMed).

Dong, Wang, Kao, Diaz, Tasset, Kaushik, Thiruthuvanathan, Zintiridou, Nieves, Dzieciatkowska, Reisz, Gavathiotis, DAlessandro, Will, Cuervo: "Chaperone-mediated autophagy sustains haematopoietic stem-cell function." in: Nature, Vol. 591, Issue 7848, pp. 117-123, (2021) (PubMed).

Chen, Li, Ni, Chen, Yang, He, Luo: "Acute brain vascular regeneration occurs via lymphatic transdifferentiation." in: Developmental cell, Vol. 56, Issue 22, pp. 3115-3127.e6, (2021) (PubMed).

Dong, Aguirre-Hernandez, Scrivo, Eliscovich, Arias, Bravo-Cordero, Cuervo: "Monitoring spatiotemporal changes in chaperone-mediated autophagy in vivo." in: Nature communications, Vol. 11, Issue 1, pp. 645, (2020) (PubMed).

Chen, He, Ni, Yang, Zhang, Luo: "Cerebrovascular Injuries Induce Lymphatic Invasion into Brain Parenchyma to Guide Vascular Regeneration in Zebrafish." in: Developmental cell, Vol. 49, Issue 5, pp. 697-710.e5, (2019) (PubMed).

Patrizio, Renner, Pizzarelli, Triller, Specht: "Alpha subunit-dependent glycine receptor clustering and regulation of synaptic receptor numbers." in: Scientific reports, Vol. 7, Issue 1, pp. 10899, (2018) (PubMed).

Bergé, Campagne, Mignolet, Holden, Théraulaz, Manley, Allain, Viollier: "Modularity and determinants of a (bi-)polarization control system from free-living and obligate intracellular bacteria." in: eLife, Vol. 5, (2017) (PubMed).

McEwen, Maher, Mo, Gottardi: "E-cadherin phosphorylation occurs during its biosynthesis to promote its cell surface stability and adhesion." in: Molecular biology of the cell, Vol. 25, Issue 16, pp. 2365-74, (2015) (PubMed).

Antigendetails

Target: Dendra 2