Invitrogen™ AIRE Monoclonal Antibody (5H12), eFluor™ 660, eBioscience™, Invitrogen™

Catalog No.	Quantity
50-112-4413	100 μg
50-112-4412	25 μg

Catalog No. 50-112-4413 Supplier Invitrogen™ Supplier No. 50593482

Description

Rat Monoclonal Antibody

Description: This 5H12 monoclonal antibody reacts with mouse Autoimmune Regulator (AIRE). This 58-kDa transcription factor is highly expressed within the perinuclear region of mature medullary thymic epithelial cells (mTECs). However, expression of AIRE protein in the periphery remains unclear despite the detection of AIRE mRNA in the lymph node, lymphoid stromal cells, monocytes, and dendritic cells. Studies demonstrate that AIRE is involved in thymocyte development, in particular the regulation of tissue-specific antigen expression during negative selection. Other biological functions for AIRE include chemokine expression and antigen presentation. The 5H12 antibody recognizes mouse AIRE isoforms 1 and 2. Applications Reported: This 5H12 antibody has been reported for use in intracellular flow cytometric analysis and immunohistochemical staining of frozen tissue sections. Applications Tested: This 5H12 antibody has been tested by immunohistochemistry on frozen 5-week-old mouse thymus and can be used at less than or equal to 10 μg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. eFluor™ 660 is a replacement for Alexa Fluor™ 647. eFluor™ 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome. Excitation: 633-647 nm; Emission: 668 nm; Laser: Red Laser. Filtration: 0.2 μm post-manufacturing filtered.

AIRE is a transcriptional regulator protein that binds to DNA as a dimer and tetramer, but not as a monomer, and may be involved in the regulation of gene expression. A nuclear protein, AIRE is a 545 amino acid protein with an HSR domain, two PHD type zinc fingers, and a SAND domain, and may be present in soluble high-molecular-weight complexes. It may form homodimer, homotetramer and also nuclear bodies and interacts with the transcriptional coactivator CBP and CREBBP. Three isoforms of AIRE are reported and one of them is predicted to be a candidate for nuclear-mediated decay (NMD). AIRE is involved in immune regulation and is known to mediate E3 ubiquitin ligase activity. Defects in AIRE are responsible for the rare autosomal-recessive systemic autoimmune disease termed autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED).

Specifications

• Antigen: AIRE
• Applications: Flow Cytometry, Immunohistochemistry (Frozen), Immunocytochemistry
• Classification: Monoclonal
• Clone: 5H12
• Concentration: 0.2 mg/mL
• Conjugate: eFluor 660
• Formulation: PBS with 0.09% sodium azide; pH 7.2
• Gene: Aire
• Gene Accession No.: Q9Z0E3
• Gene Alias: Aire; AIRE protein; AIRE1; APECED; APECED protein; APECED protein homolog; APS1; APSI; Autoimmune polyendocrinopathy candidiasis ectodermal dystrophy protein; autoimmune polyendocrinopathy candidiasis ectodermal dystrophy protein homolog; Autoimmune regulator; autoimmune regulator (autoimmune polyendocrinopathy candidiasis ectodermal dystrophy); autoimmune regulator AIRE1a; LOW QUALITY PROTEIN: autoimmune regulator; OTTHUMP00000109529; PGA1
• Gene Symbols: Aire
• Host Species: Rat
• Purification Method: Affinity chromatography
• Quantity: 100 μg
• Regulatory Status: RUO
• Primary or Secondary: Primary
• Gene ID (Entrez): 11634
• Target Species: Mouse
• Content And Storage: 4°C, store in dark, DO NOT FREEZE!
• Product Type: Antibody
• Form: Liquid
• Isotype: IgG2c κ

Frequently Asked Questions (FAQs)

I am unable to analyze my cells stained with eFluor Organic Dyes today. What options do I have?

Our options will depend on the samples you are analyzing.

If cell viability is not critical, you can store your stained samples at 4 degrees C or on ice overnight in the dark and analyze the following day.

For samples stained with eFluor organic fluorochromes, we recommend that cells be suspended in 100 uL of Flow Cytometry Staining Buffer (Cat. No. 00-4222) and 100 uL of eBioscience IC Fixation Buffer (Cat. No. 00-8222); samples can be incubated for up to 3 days at 4 degrees C in the dark. Alternatively, the 1-step Fix/Lyse Solution (Cat. No. 00-5333) can be used. This is a great option when working with whole blood but also works for other cell types.

Can the eFluor Organic fluorochromes be used for intracellular staining?

Yes, the eFluor Organic fluorochromes can be used for intracellular staining. The eFluor organic fluorochromes maintain bright signal and require minimal changes in compensation when fixed with eBioscience IC Fixation Buffer (Cat. No. 00-8222-49) and Permeabilization Buffer (Cat. No. 00-8333-56) or 1-step Fix/Lyse Solution (Cat. No. 00-5333-54, 00-5333-57) (as compared to live cells).

Is the eFluor 660 fluorochrome compatible with Anti-Cy5/Alexa Fluor 647 beads?

Yes, in-house studies have demonstrated that the eFluor 660 fluorochrome is recognized by Anti-Cy5/Alexa Fluor 647 beads. Side by side studies with Alexa Fluor 647 versus eFluor 660 conjugated antibodies have demonstrated comparable results.

What buffers are compatible with the eFluor Organic fluorochromes and eVolve QDots?

The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.

What is the difference between eFluor Organic Dyes and eVolve Qdots?

The eFluor Organic Dyes (eFluor 450, APC-eFluor 780, PerCP-eFluor 710, eFluor 710) are conventional fluorochromes. In contrast, the eVolve line of products are Quantum dots.

Are the eFluor Organic Dyes photo-labile?

As with other fluorochromes, we recommend minimal exposure to light to maintain optimal signal.

For Research Use Only.