Anti-IL-2 Receptor alpha/Il2ra Antibody Picoband™

Product Name

Anti-IL-2 Receptor alpha/Il2ra Antibody Picoband™

View all CD25/IL-2R alpha Antibodies

SKU/Catalog Number

A00214-1

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-IL-2 Receptor alpha/Il2ra Antibody Picoband™ catalog # A00214-1. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-IL-2 Receptor alpha/Il2ra Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00214-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

A synthetic peptide corresponding to a sequence in the middle region of mouse IL-2 Receptor alpha/Il2ra, which shares 64.3% and 86.7% amino acid (aa) sequence identity with human and rat IL-2 Receptor alpha/Il2ra, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

A00214-1 is reactive to IL2RA in Mouse, Rat

Applications

A00214-1 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

50 kDa

Calculated molecular weight

30.819kDa

Background of CD25/IL-2R alpha

IL2RA(Interleukin 2 receptor, alpha), also called IL2R, IL2R, Alpha Chain or CD25, is a protein that in humans is encoded by the IL2RA gene.The IL2RA gene is mapped on 10p15.1. IL2RA is composed of strand-swapped "sushi-like" domains, unlike the classical cytokine receptor fold. As a result of this domain swap, IL2RA uses a composite surface to dock into a groove on IL2 that also serves as a binding site for antagonist drugs.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. Infection by the protozoan Trypanosoma cruzi causes Chagas disease, characterized by a reduction in the amount of IL2RA expressed on the surface of immune cells. This leads to chronic immune suppression, becoming increasingly severe over the course of many years and ultimately resulting in death if left untreated.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Mouse, Rat
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Mouse, Rat
Flow Cytometry, 1-3 μg/1x10⁶ cells, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of IL-2 Receptor alpha/Il2ra using anti-IL-2 Receptor alpha/Il2ra antibody (A00214-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: mouse ANA-1 whole cell lysates,
Lane 2: mouse NIH/3T3 whole cell lysates,
Lane 3: mouse RAW264.7 whole cell lysates,
Lane 4: mouse HEPA1-6 whole cell lysates,
Lane 5: rat C6 whole cell lysates,
Lane 6: rat NRK whole cell lysates,
Lane 7: rat RH35 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-IL-2 Receptor alpha/Il2ra antigen affinity purified polyclonal antibody (Catalog # A00214-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for IL-2 Receptor alpha/Il2ra at approximately 50 kDa. The expected band size for IL-2 Receptor alpha/Il2ra is at 50 kDa.

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Figure 2. IHC analysis of IL-2 Receptor alpha/Il2ra using anti-IL-2 Receptor alpha/Il2ra antibody (A00214-1).
IL-2 Receptor alpha/Il2ra was detected in a paraffin-embedded section of mouse spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IL-2 Receptor alpha/Il2ra Antibody (A00214-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 3. IHC analysis of IL-2 Receptor alpha/Il2ra using anti-IL-2 Receptor alpha/Il2ra antibody (A00214-1).
IL-2 Receptor alpha/Il2ra was detected in a paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-IL-2 Receptor alpha/Il2ra Antibody (A00214-1) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of HEPA1-6 cells using anti-IL-2 Receptor alpha/Il2ra antibody (A00214-1).
Overlay histogram showing HEPA1-6 cells stained with A00214-1 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-IL-2 Receptor alpha/Il2ra Antibody (A00214-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For IL2RA (Source: Uniprot.org, NCBI)

Alternative Names

CD25 antigen; CD25; IDDM10; IL-2 R alpha; IL-2 receptor subunit alpha; IL2R alpha; IL-2R subunit alpha; IL2R; IL2RA; IL-2Ra; IL-2-RA; IL2-RA; interleukin 2 receptor, alpha; interleukin-2 receptor subunit alpha; p55; TAC antigen; TCGFR IL2RA CD25, IDDM10, IL2R, IMD41, TCGFR, p55 interleukin 2 receptor subunit alpha interleukin-2 receptor subunit alpha|IL-2 receptor subunit alpha|IL-2R subunit alpha|TAC antigen|interleukin 2 receptor, alpha

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