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CLNS1A antibody

CLNS1A Reactivity: Human WB, FACS, IHC Host: Rabbit Polyclonal unconjugated
Catalog No. ABIN4903315
  • Target See all CLNS1A Antibodies
    CLNS1A (Chloride Channel, Nucleotide-Sensitive, 1A (CLNS1A))
    Reactivity
    • 42
    • 21
    • 19
    • 4
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Host
    • 39
    • 3
    Rabbit
    Clonality
    • 39
    • 3
    Polyclonal
    Conjugate
    • 22
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This CLNS1A antibody is un-conjugated
    Application
    • 35
    • 13
    • 7
    • 6
    • 5
    • 4
    • 3
    • 3
    • 3
    • 2
    • 2
    • 1
    Western Blotting (WB), Flow Cytometry (FACS), Immunohistochemistry (IHC)
    Purification
    Affinity purification
    Immunogen
    A synthetic Peptide of human CLNS1A
    Isotype
    IgG
    Top Product
    Discover our top product CLNS1A Primary Antibody
  • Application Notes
    WB 1:500 - 1:1000
    IHC 1:20 - 1:50
    FC 1:20 - 1:50
    Restrictions
    For Research Use only
  • Validation #102909 (Western Blotting)
    'Independent Validation' Badge
    by
    AG Gruss, Institut für Genetik, Rheinische Friedrich-Wilhelms-Universität Bonn
    No.
    #102909
    Date
    09/20/2018
    Antigen
    CLNS1A
    Lot Number
    03R181011
    Method validated
    Western Blotting
    Positive Control

    HeLa cells

    Independent antibody

    Negative Control

    HeLa cells, CLNS1A knockdown using Dharmacon smartpool siRNA

    Notes

    Passed. ABIN4903315 specifically reveals endogenous CLNS1A in HeLa cell lysates while no signal is detected subsequently to knock-down of the antigen.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN4903315
    Secondary Antibody
    HRP-conjugated goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch, 111-035-003)
    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.
    • 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
      • rabbit anti-CLNS1A antibody (antibodies-online, ABIN4903315, lot 03R181011) 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
      • HRP-conjugated goat anti-rabbit IgG (H+L) (Jackson ImmunoResearch, 111-035-003) diluted 1:1000 in 1x Roti Block for 1h at RT,
      • goat anti-mouse IgG (H+L) (Jackson ImmunoResearch, 115-005-003, lot 128525) diluted 1:10000 in PBS for 45min at RT, or
      • HRP-conjugated goat anti-rat IgG (H+L) (Jackson ImmunoResearch, 112-035-003) diluted 1:10000 in PBS for 45min at RT.
    • Wash membrane 5x for 5 min with TBS-T.
    • Reveal protein bands using ECL (GE Healthcare, RPN3244V2, 9633722) on an LAS mini system (GE healthcare).
    Experimental 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.

  • Format
    Liquid
    Buffer
    PBS with 0.02 % sodium azide, 50 % glycerol,  pH 7.3.
    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Handling Advice
    Avoid repeated freeze/thaw cycles.
    Storage
    -20 °C
    Storage Comment
    Store at -20C. Avoid freeze / thaw cycles.
  • Target
    CLNS1A (Chloride Channel, Nucleotide-Sensitive, 1A (CLNS1A))
    Alternative Name
    CLNS1A (CLNS1A Products)
    Synonyms
    CLCI antibody, CLNS1B antibody, ICln antibody, 2610036D06Rik antibody, 2610100O04Rik antibody, Clci antibody, Clcni antibody, ICLN antibody, Icln antibody, pICln antibody, clci antibody, clns1b antibody, icln antibody, pIcIn antibody, chloride nucleotide-sensitive channel 1A antibody, chloride channel, nucleotide-sensitive, 1A antibody, chloride channel, nucleotide-sensitive, 1A L homeolog antibody, CLNS1A antibody, Clns1a antibody, clns1a.L antibody, clns1a antibody
    Background
    This gene encodes a protein that functions in multiple regulatory pathways. The encoded protein complexes with numerous cytosolic proteins and performs diverse functions including regulation of small nuclear ribonucleoprotein biosynthesis, platelet activation and cytoskeletal organization. The protein is also found associated with the plasma membrane where it functions as a chloride current regulator. Pseudogenes of this gene are found on chromosomes 1, 4 and 6. Several transcript variants encoding different isoforms have been found for this gene.
    Molecular Weight
    26.215 kDa
    Gene ID
    1207
    UniProt
    P54105
    Pathways
    Ribonucleoprotein Complex Subunit Organization
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