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CLNS1A anticorps

CLNS1A Reactivité: Humain WB, ICC, DB Hôte: Souris Monoclonal 2187C2a unconjugated
N° du produit ABIN933127
  • Antigène Voir toutes CLNS1A Anticorps
    CLNS1A (Chloride Channel, Nucleotide-Sensitive, 1A (CLNS1A))
    Reactivité
    • 34
    • 20
    • 19
    • 4
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Humain
    Hôte
    • 32
    • 2
    Souris
    Clonalité
    • 32
    • 2
    Monoclonal
    Conjugué
    • 20
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Cet anticorp CLNS1A est non-conjugé
    Application
    • 33
    • 13
    • 7
    • 6
    • 5
    • 4
    • 4
    • 3
    • 3
    • 2
    • 2
    • 1
    Western Blotting (WB), Immunocytochemistry (ICC), Dot Blot (DB)
     Réactivité croisée
    Humain, Souris, Rat (Rattus)
    Réactivité croisée (Details)
    Other species not studied.
    Purification
    Protein G affinity chromatography
    Immunogène
    CLNS1 A antibody was raised in mouse using recombinant Human Chloride Channel, Nucleotide-Sensitive, 1 (Clns1 )
    Clone
    2187C2a
    Isotype
    IgG1
    Top Product
    Discover our top product CLNS1A Anticorps primaire
  • Indications d'application
    WB: 0.2-2 µg/mL, ICC: 2-100 µg/mL
    Optimal conditions should be determined by the investigator.
    Restrictions
    For Research Use only
  • Validation #102907 (Western Blotting)
    'Independent Validation' signe
    by
    AG Gruss, Institut für Genetik, Rheinische Friedrich-Wilhelms-Universität Bonn
    No.
    #102907
    Date
    20.09.2018
    Antigène
    CLNS1A
    Numéro du lot
    9832
    Application validée
    Western Blotting
    Contrôle positif

    HeLa cells

    Independent antibody

    Contrôle négative

    HeLa cells, CLNS1A knockdown using Dharmacon smartpool siRNA

    Conclusion

    Passed. ABIN933127 specifically reveals endogenous CLNS1A in HeLa cell lysates while no signal is detected subsequently to knock-down of the antigen.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN933127
    Anticorps secondaire
    goat anti-mouse IgG (H+L) (Jackson ImmunoResearch, 115-005-003, lot 128525)
    Full Protocol
    • Seed 3x104 HeLa cells in DMEM (PAN Biotech, P04-41450, 2730618) supplemented with 10% FBS (PAN Biotech, P03-3302), without antibiotics, in 6 well plates in 2 ml media.
    • Grow for 24h cells at 37°C and 5% CO2 to 70% confluency.
    • Change medium.
    • Transfect cells with 60pmol ON-TARGETplus SmartPool siRNA human (Dharmacon, L-012571-00-0005, lot 170831) in 125µl 150mM NaCl added to 7.5µl Lipofectamine RNAiMax (13778030, lot 1911816) in 125µl 150mM NaCl.
    • Grow cells for 24h at 37°C and 5% CO2.
    • Change medium.
    • Grow cells for 72h at 37°C and 5% CO2.
    • Wash cells once with PBS (PAN Biotech, P04-36500, lot 3620618).
    • Detach cells with 100µl trypsin per well.
    • For WB the cells were once washed with PBS (PAN Biotech) before treated with 100µl of Trypsin (PAN Biotech, P10-023100, lot 6860417) per well.
    • Once cells are detached add 900µl medium.
    • Count cells.
    • Add 40µl 1x Laemmli buffer (Roth, K929.1, lot 227259243) to 2x105 cells. Heat up to 99°C for 10min.
    • Separate 10µl of the lysates on a freshly cast denaturing 12% SDS-PAGE gel in an electrophoresis chamber for 1h at 120V.
    • Transfer proteins onto PVDF membrane (Merck, IPVH0010, lot K5KN7128I) with a wet blotting system for 1h at 80V.
    • Block the membrane with 1x Roti Block (Roth, A151.2, lot 148270222) for 30min at RT.
    • Incubation with primary
      • mouse anti-CLNS1A antibody (antibodies-online, ABIN933127, lot 9832) diluted 1:500 in 1x Roti Block for 1h at RT,
      • mouse anti-CLNS1A antibody (Santa Cruz, sc-271454, G3010) diluted 1:3000 in 1x Roti Block for 1h at RT, or
      • rat anti tubulin antibody (YOL 1/34 gifted) diluted 1:1000 in 1x Roti Block for 1h at RT.
    • Wash membrane 2x for 10min with 1x TBST.
    • Incubation with secondary
      • goat anti-mouse IgG (H+L) (Jackson ImmunoResearch, 115-005-003, lot 128525) diluted 1:10000 in PBS for 45min at RT or
      • goat anti-rat IgG (H+L) (Jackson ImmunoResearch, 112-035-003) diluted 1:10000 in PBS for 45min at RT.
    • Wash membrane 5x for 5min with 1x TBS-T.
    • Reveal protein bands using ECL (GE Healthcare, RPN3244V2, 9633722) on an LAS mini system (GE healthcare).
    Notes

    Western blot using ABIN933127 on HeLa cell extracts reveals a band of the expected molecular weight. The same band is also detected by an independent CLNS1A antibody and disappears upon siRNA knock-down.

  • Validation #103452 (Immunofluorescence)
    'Independent Validation' signe
    by
    AG Gruss, Institut für Genetik, Rheinische Friedrich-Wilhelms-Universität Bonn
    No.
    #103452
    Date
    20.09.2018
    Antigène
    CLNS1A
    Numéro du lot
    9832
    Application validée
    Immunofluorescence
    Contrôle positif

    HeLa cells

    Independent antibody

    Contrôle négative

    HeLa cells, CLNS1A knockdown using Dharmacon smartpool siRNA

    Conclusion

    Passed. ABIN933127 specifically reveals endogenous CLNS1A in HeLa cells while no signal is detected subsequently to knock-down of the antigen.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN933127
    Anticorps secondaire
    AF488 conjugated secondary goat anti-mouse IgG (H+L) (ThermoFisher Scientific, A32723)
    Full Protocol
    • Seed 3x104 HeLa cells in DMEM (PAN Biotech, P04-41450, 2730618) supplemented with 10% FBS (PAN Biotech, P03-3302), without antibiotics, in 6 well plates in 2ml media.
    • Grow cells for 24h at 37°C and 5% CO2 to 70% confluency.
    • Change medium.
    • Transfect cells with 60pmol ON-TARGETplus SmartPool siRNA human (Dharmacon, L-012571-00-0005, lot 170831) in 125µl 150mM NaCl added to 7.5µl Lipofectamine RNAiMax (13778030, lot 1911816) in 125µl 150mM NaCl.
    • Grow cells for 24h at 37°C and 5% CO2.
    • Change medium.
    • Grow cells for 72h at 37°C and 5% CO2.
    • Transfer cells from each well on five coverslips.
    • Grow cells for 96h at 37°C and 5% CO2.
    • Wash cells with 1x PBS (PAN Biotech).
    • Add 4% PFA for 10min at 37°C.
    • Wash cells 2x with 1x PBS 1x.
    • Wash cells 1x 5min with 0.5% Triton X-100.
    • Wash cells 2x with 1x PBS containing 1x Roti-Immunoblock (Roth, T144.1, lot 248272501).
    • Store cells ON at 4°C.
    • Incubation with primary
      • mouse anti-CLNS1A antibody (antibodies-online, ABIN933127, lot 9832) diluted 1:500 in Roti-Immunoblock for 1h at RT or
      • mouse anti-CLNS1A antibody (Santa Cruz, sc-271454, G3010) diluted 1:500 in Roti-Immunoblock for 1h at RT.
    • Wash coverslips with PBS.
    • Incubation with AF488 conjugated secondary goat anti-mouse IgG (H+L) (ThermoFisher Scientific, A32723) diluted 1:1000 for 45min at RT.
    • Wash coverslips with PBS.
    • Mount coverslips on glass slides in Fluoromount-G mounting medium (Southern Biotech, 0100-01).
    • Image acquisition with an exposure time of 600ms (ABIN933127) or 400ms (sc-271454) on (Zeiss Axiophot, 63x magnification, resolution 1388x1040 pixels).
    Notes

    IF staining of HeLA cells using ABIN933127 reveals the expected staining pattern. Cellular staining was not observed after CLNS1A knock-down.

  • Concentration
    Lot specific
    Buffer
    CLNS1 A antibody in PBS (3.0 mM KCl, 1.5 mM KH2 PO4 , 140 mM NaCl, 8.0 mM Na2 HPO4 (pH 7.4)) containing 1 % bovine serum albumin (BSA) and 0.05 % 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
    Avoid repeated freeze/thaw cycles.
    Dilute only prior to immediate use.
    Stock
    4 °C/-20 °C
    Stockage commentaire
    Store at 2-8 °C for up to one year. We recommend long term storage at -20 °C.
  • Antigène
    CLNS1A (Chloride Channel, Nucleotide-Sensitive, 1A (CLNS1A))
    Autre désignation
    CLNS1A (CLNS1A Produits)
    Synonymes
    anticorps CLCI, anticorps CLNS1B, anticorps ICln, anticorps 2610036D06Rik, anticorps 2610100O04Rik, anticorps Clci, anticorps Clcni, anticorps ICLN, anticorps Icln, anticorps pICln, anticorps clci, anticorps clns1b, anticorps icln, anticorps pIcIn, anticorps chloride nucleotide-sensitive channel 1A, anticorps chloride channel, nucleotide-sensitive, 1A, anticorps chloride channel, nucleotide-sensitive, 1A L homeolog, anticorps CLNS1A, anticorps Clns1a, anticorps clns1a.L, anticorps clns1a
    Sujet
    The interaction with Sm proteins inhibits their assembly on U RNA and interferes with snRNP biogenesis. Inhibits the binding of survival motor neuron protein (SMN) to Sm proteins. May participate in cellular volume control by activation of a swelling-induced chloride conductance pathway. Synonyms: Monoclonal CLNS1A antibody, Anti-CLNS1A antibody, Chloride channel nucleotide sensitive 1A antibody, CLCI antibody, ICln antibody, CLNS1B antibody.
    Pathways
    Ribonucleoprotein Complex Subunit Organization
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