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CD25 antibody (AA 201-268)

IL2RA Reactivity: Human, Mouse FACS, WB, ELISA, IHC (p) Host: Rabbit Polyclonal unconjugated
Catalog No. ABIN670371
  • Target See all CD25 (IL2RA) Antibodies
    CD25 (IL2RA) (Interleukin 2 Receptor, alpha (IL2RA))
    Binding Specificity
    • 15
    • 10
    • 6
    • 6
    • 5
    • 5
    • 4
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    AA 201-268
    Reactivity
    • 170
    • 106
    • 35
    • 9
    • 6
    • 3
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    Human, Mouse
    Host
    • 129
    • 79
    • 78
    • 2
    • 1
    Rabbit
    Clonality
    • 209
    • 76
    • 2
    Polyclonal
    Conjugate
    • 105
    • 31
    • 26
    • 25
    • 19
    • 9
    • 6
    • 5
    • 4
    • 4
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This CD25 antibody is un-conjugated
    Application
    • 201
    • 84
    • 50
    • 46
    • 39
    • 33
    • 19
    • 16
    • 14
    • 14
    • 8
    • 6
    • 4
    • 4
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Flow Cytometry (FACS), Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
    Cross-Reactivity
    Human, Mouse
    Predicted Reactivity
    Rat
    Purification
    Purified by Protein A.
    Immunogen
    KLH conjugated synthetic peptide derived from mouse CD25/IL2RA
    Isotype
    IgG
  • Application Notes
    WB 1:300-5000
    ELISA 1:500-1000
    FCM 1:20-100
    IHC-P 1:200-400
    Restrictions
    For Research Use only
  • Validation #104539 (Immunohistochemistry)
    'Independent Validation' Badge
    by
    TRON
    No.
    #104539
    Date
    02/28/2024
    Antigen
    CD25
    Lot Number
    AG10271265
    Method validated
    Immunohistochemistry
    Positive Control

    FFPE human tonsil

    Negative Control
    Notes

    Passed. The CD25 antibody ABIN670371 recognizes CD25 in human tonsil tissue.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN670371
    Secondary Antibody
    HRP-conjugated goat anti-rabbit antibody (medac diagnostics, DPVR-110HRP, Lot 101122)
    Full Protocol
    • Dewax and rehydrate tissue sections on glass slides in a descending xylene and graded alcohol series:
      • xylene 2x for 5 min.
      • 100 % (v/v) ethanol 2x for 5 min.
      • 96 % (v/v) ethanol 2x for 5 min.
      • 70 % (v/v) ethanol 2x for 5 min.
    • Wash slides 2x 5 min with ddH2O.
    • Cook slides for 30 min at 95 °C in citrate buffer pH 6.
    • Wash slides 2x 3 min with ddH2O.
    • Incubate slides in 0.3 % (v/v) H2O2 in PBS for 15 min to quench endogenous peroxidase.
    • Wash slides 3x 3 min in 1X PBS.
    • Block slides in 10 % goat serum in PBS for 30 min at RT.
    • Incubate slides with primary rabbit anti-CD25 antibody (antibodies-online, ABIN670371, lot AG10271265) diluted 1:250 and 1:500 in 10 % goat serum in PBS ON at 4 °C in a humidified chamber.
    • Wash slides 3x 3 min in 1X PBS.
    • Incubate slides with secondary HRP-conjugated goat anti-rabbit antibody (medac diagnostics, DPVR-110HRP, Lot 101122) ready-to-use in 10 % goat serum in PBS for 30 min at RT in a humidified chamber.
    • Wash slides 3x 3 min in 1X PBS.
    • HRP development.
    • Counterstain slides with hematoxylin solution (Carl Roth, T865.2).
    • Wash slides for 15 min under running tap water.
    • Dehydrate sections in an ascending alcohol-xylene series:
      • 70% ethanol 2x for 10 sec.
      • 96% ethanol 2x for 10 sec.
      • 100% ethanol 2x for 10 sec.
      • xylene 2x for 10 sec.
    • Mount sections with Entellan new mounting medium (Sigma-Aldrich, 1008690500).
    • Dry sections.
    • Image acquisition on an Aperio CS2 Digital Pathology Slide Scanner (Leica), Magnification 40x.
    Experimental Notes
    • ABIN670371 appears to successfully recognize CD25 in human tonsils, but in mouse spleen and lung, the signal is not present where expected.

    • ABIN670371 does not appear to recognize CD25 in mouse samples. CD25 should be highly expressed on Tregs, which by definition express FoxP3. ABIN670371 signal does not co-localize with FoxP3 in either spleen or lung. Furthermore, in both tissues, the signal from ABIN670371 is nuclear, whereas it is expected to be membrane. ABIN670371 signal does not co-localize with CD8/CD4 in either spleen or lung although CD25 should (at least partially) co-localize with CD8 and CD4.

  • Format
    Liquid
    Concentration
    1 μg/μL
    Buffer
    0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.
    Preservative
    ProClin
    Precaution of Use
    This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
    Storage
    4 °C,-20 °C
    Storage Comment
    Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
    Expiry Date
    12 months
  • Target
    CD25 (IL2RA) (Interleukin 2 Receptor, alpha (IL2RA))
    Alternative Name
    IL2RA (IL2RA Products)
    Synonyms
    CD25 antibody, IDDM10 antibody, IL2R antibody, TCGFR antibody, IL2RAC antibody, IL-2RA antibody, Il2r antibody, Ly-43 antibody, interleukin 2 receptor subunit alpha antibody, interleukin-2 receptor subunit alpha-like antibody, interleukin 2 receptor, alpha chain antibody, IL2RA antibody, Il2ra antibody, LOC100222957 antibody
    Background

    Synonyms: CD25, Il2r, Ly-43, Interleukin-2 receptor subunit alpha, IL-2 receptor subunit alpha, IL-2-RA, IL-2R subunit alpha, IL2-RA, p55, Il2ra

    Background: The interleukin 2 (IL2) receptor alpha (IL2RA) and beta (IL2RB) chains, together with the common gamma chain (IL2RG), constitute the high-affinity IL2 receptor. Homodimeric alpha chains (IL2RA) result in low-affinity receptor, while homodimeric beta (IL2RB) chains produce a medium-affinity receptor. Normally an integral-membrane protein, soluble IL2RA has been isolated and determined to result from extracellular proteolyisis. Alternately-spliced IL2RA mRNAs have been isolated, but the significance of each is presently unknown. Mutations in this gene are associated with interleukin 2 receptor alpha deficiency.[provided by RefSeq, Nov 2009].

    Gene ID
    16184
    UniProt
    P01590
    Pathways
    JAK-STAT Signaling, Growth Factor Binding, Activated T Cell Proliferation
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