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Goat anti-Rabbit IgG (Heavy & Light Chain) Antibody (FITC) - Preadsorbed Secondary Antibody

FLISA, FACS, FM Polyclonal IgG FITC
Pubmed (23) Independent Validation (1)
Catalog No. ABIN101988
$223.50
Plus shipping costs $45.00
2 mg ABIN101988
2 mg ABIN101988
local_shipping Shipping to: United States
Delivery in 1 to 2 Business Days
  • Target
    IgG
    Binding Specificity
    • 1528
    • 134
    • 117
    • 46
    • 34
    • 30
    • 25
    • 6
    • 6
    • 6
    • 6
    • 5
    • 4
    • 4
    • 4
    • 3
    • 2
    • 1
    • 1
    Heavy & Light Chain
    Reactivity
    • 367
    • 333
    • 314
    • 224
    • 213
    • 111
    • 110
    • 82
    • 76
    • 73
    • 72
    • 65
    • 61
    • 56
    • 54
    • 49
    • 42
    • 31
    • 27
    • 15
    • 9
    • 7
    • 5
    • 4
    • 4
    • 4
    • 4
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Rabbit
    Host
    • 821
    • 781
    • 312
    • 178
    • 76
    • 75
    • 22
    • 20
    • 16
    • 12
    • 5
    • 4
    • 4
    • 3
    • 1
    • 1
    • 1
    Goat
    Clonality
    • 1786
    • 177
    Polyclonal
    Conjugate
    • 317
    • 233
    • 215
    • 207
    • 147
    • 146
    • 128
    • 72
    • 49
    • 46
    • 36
    • 31
    • 29
    • 28
    • 26
    • 26
    • 26
    • 25
    • 25
    • 25
    • 24
    • 24
    • 23
    • 23
    • 23
    • 23
    • 23
    • 22
    • 21
    • 19
    • 19
    • 18
    • 18
    • 14
    • 12
    • 11
    • 10
    • 9
    • 8
    • 8
    • 6
    • 6
    • 6
    • 6
    • 5
    • 5
    • 5
    • 5
    • 5
    • 5
    • 5
    • 5
    • 4
    • 4
    • 4
    • 4
    • 4
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    FITC
    Application
    • 1261
    • 1110
    • 917
    • 807
    • 539
    • 524
    • 498
    • 144
    • 100
    • 71
    • 71
    • 65
    • 52
    • 50
    • 29
    • 28
    • 16
    • 13
    • 11
    • 10
    • 10
    • 9
    • 4
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    FLISA, Flow Cytometry (FACS), Fluorescence Microscopy (FM)
    Specificity
    IgG (H&L)
    Cross-Reactivity
    Rabbit
    Characteristics
    Concentration Definition: by UV absorbance at 280 nm
    Purification
    Preadsorption: Solid phase absorption
    Immunogen

    Immunogen: Rabbit IgG whole molecule

    Isotype
    IgG
    Labeling Ratio
    3.1
  • Application Notes

    Application Note: This product is designed for immunofluorescence microscopy, fluorescence based plate assays (FLISA) and fluorescent western blotting. This product is also suitable for multiplex analysis, including multicolor imaging, utilizing various commercial platforms.

    FLISA Dilution: 1:10,000 - 1:50,000

    Flow Cytometry Dilution: 1:500 - 1:2,500

    IF Microscopy Dilution: 1:1,000 - 1:5,000

    Comment

    Excitation/Emission wavelength: 494 nm/514 nm

    Restrictions
    For Research Use only
  • Validation #100071 (Immunofluorescence)
    'Independent Validation' Badge
    by
    Okeanos Research Laboratory, Department of Biological Sciences, Clemson University
    No.
    #100071
    Date
    09/14/2016
    Antigen
    Rabbit IgG (Heavy & Light Chain)
    Lot Number
    611-1202
    Method validated
    Immunofluorescence
    Positive Control
    Lab stock CBD-SNAP antibody
    Negative Control
    No SNAP-tag antibody
    Notes
    We validate the specificity of the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) ABIN101988 for rabbit IgG antibody.
    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN1573927
    Secondary Antibody
    ABIN101988
    Full Protocol
    • Oyster visceral mass tissue is dissected and fixed in 4% paraformaldehyde in seawater overnight.
    • Serial dehydration process using an automated ASP300S Enclosed Tissue Processor (Leica Biosystems) as follows:
      • 70% ethanol for 45min
      • 90% ethanol for 45min
      • 90% ethanol for 45min
      • 100% ethanol twice for 45min
      • xylene twice for 45min
      • paraffin wax at 58°C 3 times for 30 min
    • Tissue is mounted in a paraffin block and hardened overnight before.
    • 8µm tissue sections are retrieved from the block and collected on circular glass cover slips.
    • Heat cover slips at 60°C for 1h.
    • Deparaffination and rehydration:
      • Xylene twice for 15min
      • 100% ethanol twice for 10min
      • 95% ethanol for 10 min
      • 85% ethanol for 10 min
      • 70% ethanol for 10 min
      • 50% ethanol for 10 min
      • 30% ethanol for 10 min
      • distilled water for 10 min
      • PBS for 10 min
    • Wash tissue sections with PBS with 0.05% triton X twice for 30min.
    • Permeabilize in PBS with 0.05% triton X overnight.
    • Treatment of the tissue sections with 1mg/mL sodium borohydride in PBS three times for 5min to reduce autofluorescence.
    • Wash sections in PBS 3 times for 15 min for at RT.
    • Block sections in PBST with 1% BSA for 2 hours at RT.
    • Incubate sections with CBD-SNAP antibody (lab stock) diluted 1:200 in PBST with 1% BSA overnight at 4°C to detect the location of chitin.
    • Wash sections in PBS 3 times for 15min with PBS at RT.
    • Additionally, incubate the CBD-SNAP and SNAP-tag double-stained sections with rabbit anti-SNAP antibody (antibodies-online, ABIN1573927, lot 13D000621) diluted 1:200 in PBST with 1% BSA overnight at 4°C.
    • Wash sections in PBS 3 times for 15min with PBS at RT.
    • Incubate sections with the secondary goat anti-rabbit IgG (heavy & light chain) antibody (FITC) (antibodies-online, ABIN101988, lot 611-1202) diluted 1:400 in PBST with 1% BSA for 2h at °C.
    • Wash sections in PBS three times for 15min at RT.
    • Counterstain with 0.1µg/mL DAPI in PBS for 15min at RT.
    • Wash sections in PBS three times for 15min at RT.
    • Mount sections on a microscopic slide using 50% glycerol in PBS.
    • Seal cover slips with nail polish.
    • Confocal imaging on Leica SPE.
    • Visualization of the data performed on LAS 3D software.
    Experimental Notes
    To validate the specificity of the anti-rabbit FITC secondary antibody ABIN101988, 8µm paraffin sections of oyster visceral mass were observed in this study. We compared the fluorescence signals with immunofluorescence study. The negative control specimen was always compared with the test specimen or the positive control specimen on the same day, using the same laser power, gain, offset, accumulation/averaging settings on the Leica SPE confocal microscope. Visualization of the data was performed on LAS 3D software, with the same visualization setting to compare signal brightness. We found that the samples treated with anti-rabbit FITC secondary antibody ABIN101988 had similar fluorescence signals as the positive control Anti Rabbit Alexa 488. Excitation at the same laser wavelength and power did not generate fluorescence in the negative control section, when anti-rabbit FITC secondary antibody was applied in the absence of rabbit produced anti-SNAP antibody.
  • Format
    Lyophilized
    Reconstitution

    Reconstitution Volume: 1.0 mL

    Reconstitution Buffer: Restore with deionized water (or equivalent)

    Concentration
    2.0 mg/mL
    Buffer

    Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

    Stabilizer: 10 mg/mL Bovine Serum Albumin (BSA) - Immunoglobulin and Protease free

    Preservative: 0.01 % (w/v) Sodium Azide

    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
    Product is photosensitive and should be protected from light.
    Storage
    RT,4 °C,-20 °C
    Expiry Date
    12 months
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  • Target
    IgG
    Background

    Synonyms: Goat anti-Rabbit IgG Antibody fluorescein Conjugation, Goat anti-Rabbit IgG FITC Conjugated Antibody

    Background: Anti-Rabbit IgG Antibody Fluorescein generated in goat detects rabbit IgG. Secreted as part of the adaptive immune response by plasma B cells, immunoglobulin G constitutes 75 % of serum immunoglobulins. Immunoglobulin G binds to viruses, bacteria, as well as fungi and facilitates their destruction or neutralization via agglutination (and thereby immobilizing them), activation of the compliment cascade, and opsinization for phagocytosis. The whole IgG molecule possesses both the F(c) region, recognized by high-affinity Fc receptor proteins, as well as the F(ab) region possessing the epitope-recognition site. Both heavy and light chains of the antibody molecule are present. Secondary Antibodies are available in a variety of formats and conjugate types. When choosing a secondary antibody product, consideration must be given to species and immunoglobulin specificity, conjugate type, fragment and chain specificity, level of cross-reactivity, and host-species source and fragment composition. This Anti-Rabbit IgG (H&L) is conjugated to Fluorescein.

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