Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Knockout Validated, Western Blot, Neutralization, Flow Cytometry, CyTOF-ready

Cited:

Western Blot, Flow Cytometry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all CD25/IL-2R alpha Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human CD25/IL-2 Ra 
Glu22-Cys213
Accession # P01589

Specificity

Detects human CD25/IL‑2 R alpha in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 µg of the antibody by the LAL method.

Scientific Data Images for Human CD25/IL-2R alpha Antibody

Detection of Human CD25/IL-2R alpha antibody by Western Blot.

Detection of Human CD25/IL-2R alpha by Western Blot.

Western blot shows lysates of HDLM-2 human Hodgkin's lymphoma cell line and PC-3 human prostate cancer cell line (negative control). PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CD25/IL-2R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-223-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for CD25/IL-2R alpha at approximately 60 kDa (as indicated). GAPDH (AF5718) is shown as a loading control.This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western Blot Shows Human CD25/IL-2R alpha Specificity by Using Knockout Cell Line.

Western blot shows lysates of HDLM‑2 human Hodgkin’s lymphoma cell line and human CD25/IL-2 R alpha knockout HDLM‑2 human Hodgkin’s lymphoma cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human CD25/IL-2R alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-223-NA) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). A specific band was detected for CD25/IL-2R alpha at approximately 60 kDa (as indicated) in the parental HDLM‑2 human Hodgkin’s lymphoma cell line cell line, but is not detectable in knockout HDLM‑2 human Hodgkin’s lymphoma cell line cell line. GAPDH (AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Western Blot Buffer Group 1.

Cell Proliferation Induced by IL‑2 and Neutralization by Human CD25/IL‑2 R alpha Antibody.

Cell Proliferation Induced by IL‑2 and Neutralization by Human CD25/IL‑2 R alpha Antibody.

Recombinant Human IL-2 (202-IL) stimulates proliferation in the N1186 human T cell line in a dose-dependent manner (orange line). Proliferation elicited by Recombinant Human IL-2 (1 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human CD25/IL-2 Ra Antigen Affinity-purified Polyclonal Antibody (Catalog # AF-223-NA). The ND50 is typically 0.2-1 µg/mL.

Applications for Human CD25/IL-2R alpha Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: Human whole blood lymphocytes

Knockout Validated

CD25/IL-2R alpha is specifically detected in HDLM‑2 human Hodgkin's lymphoma cell line parental cell line but is not detectable in CD25/IL-2R alpha knockout HDLM‑2 human Hodgkin's lymphoma cell line cell line.

Western Blot

1 µg/mL
Sample: HDLM‑2 human Hodgkin's lymphoma cell line

Neutralization

Measured by its ability to neutralize IL‑2-induced proliferation in the N1186 human T cell line. The Neutralization Dose (ND50) is typically 0.2-1 µg/mL in the presence of 1 ng/mL Recombinant Human IL‑2.

Reviewed Applications

Read 3 reviews rated 4.7 using AF-223-NA in the following applications:

Flow Cytometry Panel Builder

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at -20 to -70 °C. Use a manual defrost freezer and avoid repeated freeze-thaw cycles. Do not use past expiration date.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: CD25/IL-2R alpha

Human CD25, also known as IL-2 receptor alpha (IL-2 R alpha ) and asTac antigen, was initially identified as a 55 kDa membrane glycoprotein that is capable of binding IL‑2. The IL-2 R alpha cDNA encodes a 272 amino acid residue precursor Type I membrane protein with a 21 residue signal peptide, a 219 residue extracellular region, a 19 residue transmembrane region and a 13 residue cytoplasmic domain. IL-2 R alpha lacks structural features characteristic of members of the cytokine receptor superfamily. By itself, IL-2 R alpha binds IL-2 with low affinity. However, when IL-2 R alpha is associated with the IL-2 receptor beta and gamma chains, a high affinity heterotrimeric receptor complex that transduces IL-2 signals is formed. Soluble forms of many cytokine receptors have been reported, and a soluble form of IL-2 R alpha (IL-2 sR alpha ) appears in serum, concomitant with its increased expression on cells. The function of the soluble IL-2 R alpha is unclear. Increased levels of IL-2 sR alpha in biological fluids reportedly correlate with increased T and B cell activation and immune system activation. Increased serum concentration of IL-2 sR alpha has been observed in patients with a variety of inflammatory conditions and in the course of some leukemias and lymphomas.

Long Name

Interleukin 2 Receptor alpha

Alternate Names

CD25, IL-2 R alpha, IL-2Ra, IL2R alpha, IL2RA

Entrez Gene IDs

3559 (Human); 16184 (Mouse); 25704 (Rat); 403870 (Canine); 102123605 (Cynomolgus Monkey)

Gene Symbol

IL2RA

UniProt

P01589

Additional CD25/IL-2R alpha Products

Product Documents for Human CD25/IL-2R alpha Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human CD25/IL-2R alpha Antibody

For research use only

Citations for Human CD25/IL-2R alpha Antibody

Customer Reviews for Human CD25/IL-2R alpha Antibody (3)

4.7 out of 5
3 Customer Ratings
5 Stars
67%
4 Stars
33%
3 Stars
0%
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1 Stars
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Customer Images

Showing  1 - 3 of 3 reviews Showing All
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  • Human CD25/IL-2R alpha Antibody
    Name: Katie Myers
    Application: ELISA
    Sample Tested: Serum
    Species: Primate
    Verified Customer | Posted 11/28/2022
    Used as capture antibody in NHP ELISA
    Human CD25/IL-2R alpha Antibody AF-223-NA
  • Human CD25/IL-2R alpha Antibody
    Name: Anonymous
    Application: ELISA
    Sample Tested: EDTA Plasma
    Species: Human
    Verified Customer | Posted 07/23/2019
    Human CD25/IL-2R alpha Antibody AF-223-NA
  • Human CD25/IL-2 R alpha Antibody
    Name: Chris Lyman
    Application: ELISA
    Sample Tested: Serum and Plasma
    Species: Human
    Verified Customer | Posted 11/03/2017
    This antibody served as both a capture and detection antibody in a sandwich ELISA developed to measure human serum and plasma samples.
    Human CD25/IL-2R alpha Antibody AF-223-NA

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