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Androgen Receptor antibody

AR Reactivity: Human, Mouse, Rat WB, IHC, IF, ICC Host: Rabbit Polyclonal unconjugated
Catalog No. ABIN1871059
  • Target See all Androgen Receptor (AR) Antibodies
    Androgen Receptor (AR)
    Reactivity
    • 309
    • 94
    • 84
    • 29
    • 17
    • 17
    • 15
    • 12
    • 8
    • 4
    • 4
    • 3
    • 1
    Human, Mouse, Rat
    Host
    • 257
    • 54
    • 1
    • 1
    • 1
    Rabbit
    Clonality
    • 248
    • 64
    Polyclonal
    Conjugate
    • 118
    • 26
    • 14
    • 14
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 7
    • 5
    • 5
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    This Androgen Receptor antibody is un-conjugated
    Application
    • 183
    • 92
    • 65
    • 56
    • 53
    • 37
    • 29
    • 20
    • 18
    • 17
    • 11
    • 9
    • 4
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC)
    Characteristics
    Expected molecular weight of target: 919-920 kDa
    Purification
    Affinity purification
    Immunogen
    A synthetic peptide of human AR
    Isotype
    IgG
  • Application Notes
    WB 1:500-1:2000, IHC 1:50-1:200, ICC 1:50-1:200, IF 1:50-1:200
    Restrictions
    For Research Use only
  • Validation #101745 (Immunofluorescence)
    'Independent Validation' Badge
    by
    School of Science and Engineering, Tulane University
    No.
    #101745
    Date
    12/21/2017
    Antigen
    Androgen Receptor
    Lot Number
    09Q266222
    Method validated
    Immunofluorescence
    Positive Control
    prostate from inducible ARlacZ mouse
    Negative Control
    secondary antibody only
    Notes

    Passed. Despite some weak non-specific staining, this antibody robustly labels the AR at a dilution of 1:100.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1871059
    Secondary Antibody
    goat anti-rabbit Cy3 conjugated antibody (Jackson ImmunoResearch)
    Full Protocol
    • Prostate Tissue from ARlacz transgenic mouse or wild type mouse is fixed via transcardial perfusion with 10% formalin and postfixed at 4°C overnight. Tissue from the transgenic mouse is used for a positive control while tissue from the wild-type mouse is used with the AR antibody.
    • Dewaxing and rehydration:
      • CitriSolv (Decon Laboratories, 1601H, 11068) 2x for 5min.
      • 100% ethanol 2x for 3min.
      • 95% ethanol 2x for 3min.
      • 70% ethanol 2x for 3min.
      • Rinse well in distilled H2O.
    • HIER:
      • Dilute 10x citrate buffer pH6.0 1:10 with dH2O and place in cases.
      • Place one slide into each case in the preheated pressure cooker (Hamilton Beach).
      • Antigen unmasking for 40min at 92-99°C.
      • Remove slides from pressure cooker (but leave them in the cases with citrate buffer). Let slides cool down for 20min to approximately 30°C.
    • Wash sections 2x for 5min with PBST rocking in a coplin jar.
    • Wash sections for 5min with PBS rocking in a coplin jar.
    • Permeabilize sections 10min in PBS + 0.2 Triton X-100 rocking in a coplin jar.
    • Wash sections 3x for 5min with PBST rocking in a coplin jar.
    • Aspirate liquid, circle tissue with a PAP pen and let dry.
    • Block each section with 200µl blocking solution (PBS containing 1% BSA, 5% normal goat serum, and 0.02% Triton X-100) for 1h at RT.
    • Aspirate blocking solution.
    • Incubate sections with 100µl primary
      • rabbit anti-androgen receptor antibody (antibodies-online, ABIN1871059, 09Q266222) diluted 1:50, 1:100, 1:200, and 1:250 in 1:10 blocking solution ON at 4°C.
      • rabbit anti-β-galactosidase antibody (Life Technologies, A1132, lot 1720092) diluted 1:500 in 1:10 blocking solution (in PBS) ON at 4°C.
      • Include a no primary antibody negative controls.
    • Aspirate the liquid.
    • Wash sections 2x for 5min with PBST.
    • Wash sections for 5min with PBST.
    • Incubate sections in the dark with secondary goat anti-rabbit Cy3 conjugated antibody (Jackson ImmunoResearch) diluted 1:400 in PBS for 1h at RT.
    • Aspirate the liquid in the dark.
    • Wash sections in the dark 2x for 5min with PBST.
    • Wash sections in the dark for 5min with PBST.
    • Nuclear counterstaining with 14.3mM DAPI diluted 1:50000 in PBS for 5min at RT in the dark.
    • Wash sections in the dark 2x for 5min with PBST.
    • Mount sections in mounting medium (Invitrogen, P36930, lot 1811419) put 1 drop of medium on each section and cover with a coverslip in the dark; seal your slides with nail polish to avoid dehydration. Store sections in the dark at 4°C.
    • Visualization on a microscope (Nikon, Eclipse Ti) with 20x objective; exposure time TRITC AR 100ms, DAPI 100ms.
    Experimental Notes
    • ABIN1871059 shows robust staining for the AR at all dilutions tried (A, D, E, and F). Dilution series was determined by manufacturer's recommendations.

    • The staining for the AR is clearly nuclear. This is shown by colocalization with the nuclear stain DAPI (B and C). Additionally, the staining pattern seen with the AR antibody is similar to the staining pattern of β-galactosidase seen in the ARlacz transgenic mouse. The ARlacz transgenic mouse expresses β-galactosidase (β-gal) under the control of the AR promoter. Therefore, β-gal is expressed wherever the endogenous AR is expressed.

    • Exclusion of the primary antibody eliminates nuclear staining, demonstrating that the AR staining is specific. Panels G and H show a prostate that was exposed to secondary antibody, but not primary antibody. The exposure time in Panel G is the same as that for the images shown in the previous figures. The exposure time in Panel H is 5x that shown in the previous figures. Although this isn’t the best negative control to use, we do not have prostate or global AR knockout mice to use for a control.

  • Format
    Liquid
    Concentration
    1 mg/mL
    Buffer
    PBS with 0.02 % sodium azide, 50 % glycerol, pH 7.4
    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 -20°C.
  • Target
    Androgen Receptor (AR)
    Alternative Name
    AR (AR Products)
    Synonyms
    AIS antibody, DHTR antibody, HUMARA antibody, HYSP1 antibody, KD antibody, NR3C4 antibody, SBMA antibody, SMAX1 antibody, TFM antibody, AW320017 antibody, Tfm antibody, Andr antibody, AR antibody, androgen receptor antibody, AR antibody, Ar antibody, ar antibody
    Molecular Weight
    110 kDa
    Gene ID
    367
    UniProt
    P10275
    Pathways
    Nuclear Receptor Transcription Pathway, Intracellular Steroid Hormone Receptor Signaling Pathway, Steroid Hormone Mediated Signaling Pathway, Regulation of Intracellular Steroid Hormone Receptor Signaling, Nuclear Hormone Receptor Binding, Chromatin Binding
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