CD25 anticorps (AA 201-268)

N° du produit ABIN670371

Cet anticorps anti-CD25 est un anticorps Lapin Polyclonal détectant CD25 dans FACS, WB, ELISA et IHC (p). Adapté pour Humain et Souris. Validé indépendamment pour une utilisation dans Immunohistochemistry.

Aperçu rapide pour CD25 anticorps (AA 201-268) (ABIN670371)

Antigène: CD25 (IL2RA) (Interleukin 2 Receptor, alpha (IL2RA))

Reactivité: Humain, Souris

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp CD25 est non-conjugé

Application: Flow Cytometry (FACS), Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))

Détail du produit anti-CD25 anticorps

Épitope: AA 201-268

Réactivité croisée: Humain, Souris

Homologie: Rat

Purification: Purified by Protein A.

Immunogène: KLH conjugated synthetic peptide derived from mouse CD25/IL2RA

Isotype: IgG

Information d'application

Indications d'application: WB 1:300-5000
ELISA 1:500-1000
FCM 1:20-100
IHC-P 1:200-400

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #104539 (Immunohistochemistry)

by: TRON

No.: #104539

Date: 28.02.2024

Antigène: CD25

Numéro du lot: AG10271265

Application validée: Immunohistochemistry

Contrôle positif:

FFPE human tonsil

Conclusion:

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

Validation Images

IIHC staining of human tonsil with anti-CD25 antibody ABIN670371 (1:250) and counterstain with hematoxylin.

IHC staining of human tonsil with anti-CD25 antibody ABIN670371 (1:500) and counterstain with hematoxylin.

Protocole

Anticorps primaire: ABIN670371

Anticorps secondaire: 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.

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.

Stockage

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Agent conservateur: ProClin

Précaution d'utilisation: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Stock: 4 °C,-20 °C

Stockage commentaire: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Date de péremption: 12 months

Détails sur CD25

Antigène: CD25 (IL2RA) (Interleukin 2 Receptor, alpha (IL2RA))

Autre désignation: IL2RA

Sujet:

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].

ID gène: 16184

UniProt: P01590

Pathways: Signalistation JAK/STAT, Growth Factor Binding, Activated T Cell Proliferation