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BrdU anticorps

BrdU FACS, IHC (p), ICC, IHC (fro) Hôte: Souris Monoclonal Bu20a unconjugated
N° du produit ABIN2477713
  • Antigène Voir toutes BrdU Anticorps
    BrdU (Bromodeoxyuridine (BrdU))
    Reactivité
    Veuillez nous consulter SVP
    Hôte
    • 51
    • 23
    • 9
    • 2
    Souris
    Clonalité
    • 59
    • 26
    Monoclonal
    Conjugué
    • 36
    • 7
    • 6
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Cet anticorp BrdU est non-conjugé
    Application
    • 27
    • 27
    • 24
    • 24
    • 19
    • 19
    • 14
    • 12
    • 12
    • 7
    • 7
    • 7
    • 6
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Flow Cytometry (FACS), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro))
    Attributs du produit
    Mouse anti BrdU antibody, clone Bu20a, recognizes BrdU and other thymidine analogs; 5′-chloro-2′-deoxyuridine (CldU), 5′-iodo-2′-deoxyuridine (IdU) and 2′-deoxy-5-ethynyluridine (EdU), but only shows minimal reactivity with thymidine itself
    Purification
    Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
    Immunogène
    Bromodeoxyuridine conjugated to BSA
    Clone
    Bu20a
    Isotype
    IgG1
  • Indications d'application
    Flow: 1/25-1/100, ICC 1/25-1/100
    For Flow Cytometry use 10 μl of the suggested working dilution to label 1x106 cells in 100 μl
    Restrictions
    For Research Use only
  • Validation #103189 (Immunohistochemistry)
    'Independent Validation' signe
    by
    Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin
    No.
    #103189
    Date
    05.10.2018
    Antigène
    BrdU
    Numéro du lot
    1710
    Application validée
    Immunohistochemistry
    Contrôle positif

    Adult (24 months) mouse brain and intestine (duodenum)

    Contrôle négative

    We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

    Conclusion

    Passed. ABIN2477713 works well in IHC-P, but with higher concentrations than the ones suggested by the manufacturer.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN2477713
    Anticorps secondaire
    goat anti-mouse AF488 conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237)
    Full Protocol
    • Inject mouse intraperitoneally at the dose of 0.1 mg/g body weight with BrdU (Sigma, B5002) dissolved in sterile distilled water at final concentration of 100mg/ml.
    • Perfuse mouse was perfused with paraformaldehyde 4% in 0.1M phosphate buffer (PB) pH 7.4 and post-fix samples in the same fixative for an additional 2h at RT.
    • Wash, dehydrate, and embed samples in paraffin wax.
    • Following several washes in PBS, cut intestines and brain with a microtome (6µm-thick sections) and mount sections on glass slides.
    • After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer (pH6.0).
    • Let sections cool for 20min.
    • Wash sections 2x 5min with distilled water.
    • Wash sections 5min with PBS.
    • Incubate sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) for 1h at RT to block non-specific binding sites.
    • Incubate sections with primary antibody mouse anti-BrdU (antibodies-online, ABIN2477713, lot 1710) diluted 1:10 in PBS-BSA-PLL ON at RT.
    • Wash sections 3x 5min with 0.01M PBS.
    • Incubate sections with the secondary goat anti-mouse AF488 conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237) diluted 1:200 in 0.1M PBS for 1h at RT.
    • Wash sections 3x 5min with 0.01M PBS.
    • Mount sections in Fluoroshield (Sigma, F6182, lot MKCB0153V).
    • Acquire images with Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x and 40x. Parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) were maintained unchanged for all images.
    Notes

    For the incubation with the primary antibody ABIN2477713 1:25, 1:20, 1:15 dilutions were also tested with also 0.01M PBS - 5% Normal Goat Serum (NGS Sigma, G9023, lot SLBV1396) and 0.1% Triton-X -100 as the blocking solution, but 1:10 and PBS containing 1% albumin from chicken egg white and 0.3% Triton-X gave the best results.

  • Validation #103975 (Immunohistochemistry)
    'Independent Validation' signe
    by
    Prof. Merighi, Laboratory of Neurobiology, Department of Veterinary Sciences, University of Turin
    No.
    #103975
    Date
    05.07.2018
    Antigène
    BrdU
    Numéro du lot
    1807
    Application validée
    Immunohistochemistry
    Contrôle positif

    Adult (24 months) mouse brain and intestine (duodenum)

    Contrôle négative

    We incubated slices overnight with the blocking solution only and then processed them with the secondary antibody.

    Conclusion

    Passed. ABIN2477713 works well in IHC-P on murine intestinal samples at a 1:10 dilution.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN2477713
    Anticorps secondaire
    goat anti-mouse AF488 conjugated antibody (Thermo Fisher Scientific, A11029, lot 1942237)
    Full Protocol
    • Inject mouse intraperitoneally at the dose of 0.1mg/g body weight with BrdU (Sigma, B5002) dissolved in sterile distilled water at final concentration of 100mg/ml.
    • Perfuse mouse with 4% paraformaldehyde in 0.1M phosphate buffer (PB) pH7.4 and post-fix samples in the same fixative for an additional 2h at RT.
    • Wash, dehydrate, and embed samples in paraffin wax.
    • Following several washes in PBS, cut intestines and brain with a microtome (6µm-thick sections) and mount sections on glass slides.
    • After paraffin removal, process sections by microwave antigen retrieval for 10min (95-100°C) in 10mM sodium citrate buffer pH6.0.
    • Let sections cool for 20min.
    • Wash sections 2x 5min with distilled water.
    • Wash sections 5min with PBS.
    • Block sections in PBS containing 1% albumin from chicken egg white (Sigma, A5378) and 0.3% Triton-X-100 (BioRad, 161-0407, lot 00583) for 1h at RT to block non-specific binding sites.
    • Incubate sections with primary mouse anti-BrdU antibody (antibodies-online, ABIN2477713, lot 1807) diluted 1:10 in PBS-BSA-PLL ON at RT.
    • Wash sections 3x 5min with 0.01M PBS.
    • Incubate sections with secondary goat anti-mouse AF488-conjugated antibody (Invitrogen, A11029, lot 1942237) for 1h at RT.
    • Wash sections 3x 5min with 0.01M PBS.
    • Mount sections in Fluoroshield (Sigma, F6182, lot MKCB0153V).
    • Acquire images with Leica DM 6000B fluorescence microscope equipped with the manufacturer’s FITC filter set and digital camera at magnifications of 20x and 40x. Maintain parameters for image acquisition (Exposure 2.2s, Gain 2.3x, Saturation 1.5, Gamma 1.10) unchanged for all images.
    Notes

    For the incubation with the primary antibody ABIN2477713 dilutions 1:25, 1:20, and 1:15 were also tested, but 1:10 gave the best results.

  • Format
    Liquid
    Concentration
    1.0 mg/mL
    Buffer
    Phosphate buffered saline, 0.09%Sodium Azide (NaN3)
    Agent conservateur
    Sodium azide
    Précaution d'utilisation
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
    Conseil sur la manipulation
    This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    Stock
    4 °C/-20 °C
    Stockage commentaire
    Store at +4oC or at -20oC if preferred. Storage in frost-free freezers is not recommended.
    Date de péremption
    18 months
  • Innis, Dai, Wu, Buchan, Jacobson: "Perinatal lipid nutrition alters early intestinal development and programs the response to experimental colitis in young adult rats." dans: American journal of physiology. Gastrointestinal and liver physiology, Vol. 299, Issue 6, pp. G1376-85, (2010) (PubMed).

  • Antigène
    BrdU (Bromodeoxyuridine (BrdU))
    Autre désignation
    BrdU (BrdU Produits)
    Classe de substances
    Chemical
    Sujet
    Synonyms: 5-BROMODEOXYURIDINE
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