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Western Blot (WB) (Figure 1. Western blot analysis of AIRE using anti- AIRE antibody (MBS178480). 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 50ug of sample under reducing conditions. Lane 1: rat testis tissue lysates, Lane 2: HELA whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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- AIRE antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for AIRE at approximately 58KD. The expected band size for AIRE is at 58KD. )

Rabbit AIRE Polyclonal Antibody | anti-AIRE antibody

Anti-AIRE Antibody

Gene Names
AIRE; APS1; APSI; PGA1; AIRE1; APECED
Reactivity
Human, Rat
No cross reactivity with other proteins.
Applications
Western Blot
Purity
Immunogen affinity purified.
Synonyms
AIRE; Polyclonal Antibody; Anti-AIRE Antibody; AIRE1; APECED; APECED protein; APS1; APS 1; APSI; Autoimmune regulator; PGA1; O43918; autoimmune regulator; anti-AIRE antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Human, Rat
No cross reactivity with other proteins.
Clonality
Polyclonal
Isotype
IgG
Purity/Purification
Immunogen affinity purified.
Form/Format
Lyophilized
Sequence Length
545
Applicable Applications for anti-AIRE antibody
Western Blot (WB)
Application Notes
Western Blot: 0.1-0.5mug/ml; Tested Species: Human, Rat
Tested Species:In-house tested species with positive results.
Other applications have not been tested.
Immunogen
E Coli-derived human AIRE recombinant protein (Position: M1-K120). Human AIRE shares 87.2% amino acid (aa) sequence identity with mouse AIRE.
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Preparation and Storage
At -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Western Blot (WB)

(Figure 1. Western blot analysis of AIRE using anti- AIRE antibody (MBS178480). 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 50ug of sample under reducing conditions. Lane 1: rat testis tissue lysates, Lane 2: HELA whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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- AIRE antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for AIRE at approximately 58KD. The expected band size for AIRE is at 58KD. )

Western Blot (WB) (Figure 1. Western blot analysis of AIRE using anti- AIRE antibody (MBS178480). 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 50ug of sample under reducing conditions. Lane 1: rat testis tissue lysates, Lane 2: HELA whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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- AIRE antigen affinity purified polyclonal antibody at 0.5ug/mL overnight at 4 degree 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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for AIRE at approximately 58KD. The expected band size for AIRE is at 58KD. )

Flow Cytometry (FC/FACS)

(Figure 2. Flow Cytometry analysis of HL-60 cells using anti-AIRE antibody (MBS178480).Overlay histogram showing HL-60 cells stained with MBS178480 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AIRE Antibody (MBS178480,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 2. Flow Cytometry analysis of HL-60 cells using anti-AIRE antibody (MBS178480).Overlay histogram showing HL-60 cells stained with MBS178480 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AIRE Antibody (MBS178480,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS)

(Figure 3. Flow Cytometry analysis of THP-1 cells using anti-AIRE antibody (MBS178480).Overlay histogram showing THP-1 cells stained with MBS178480 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AIRE Antibody (MBS178480,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )

Flow Cytometry (FC/FACS) (Figure 3. Flow Cytometry analysis of THP-1 cells using anti-AIRE antibody (MBS178480).Overlay histogram showing THP-1 cells stained with MBS178480 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AIRE Antibody (MBS178480,1ug/1x10^6 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10ug/1x10^6 cells) was used as secondary antibody for 30 minutes at 20 degree C. Isotype control antibody (Green line) was rabbit IgG (1ug/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control. )
Related Product Information for anti-AIRE antibody
Rabbit IgG polyclonal antibody for Autoimmune regulator (AIRE) detection.
Background: The autoimmune regulator (AIRE) is a protein that in humans is encoded by the AIRE gene. This gene encodes a transcriptional regulator that forms nuclear bodies and interacts with the transcriptional coactivator CREB binding protein. The encoded protein plays an important role in immunity by regulating the expression of autoantigens and negative selection of autoreactive T-cells in the thymus. Mutations in this gene cause the rare autosomal-recessive systemic autoimmune disease termed autoimmune polyendocrinopathy with candidiasis and ectodermal dystrophy (APECED).
References
1. Lindmark E, Chen Y, Georgoudaki AM, Dudziak D, Lindh E, Adams WC, Loré K, Winqvist O, Chambers BJ, Karlsson MC, et al. (2013). "AIRE expressing marginal zone dendritic cells balances adaptive immunity and T-follicular helper cell recruitment.". Journal of Autoimmunity 42: 62-70.
2. Liston A, Lesage S, Wilson J, Peltonen L, Goodnow CC (April 2003). "Aire regulates negative selection of organ-specific T cells". Nat. Immunol. 4(4): 350-4.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
326
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
36,501 Da
NCBI Official Full Name
autoimmune regulator
NCBI Official Synonym Full Names
autoimmune regulator
NCBI Official Symbol
AIRE
NCBI Official Synonym Symbols
APS1; APSI; PGA1; AIRE1; APECED
NCBI Protein Information
autoimmune regulator
UniProt Protein Name
Autoimmune regulator
Protein Family
UniProt Gene Name
AIRE
UniProt Synonym Gene Names
APECED; APECED protein
UniProt Entry Name
AIRE_HUMAN

NCBI Description

This gene encodes a transcriptional regulator that forms nuclear bodies and interacts with the transcriptional coactivator CREB binding protein. The encoded protein plays an important role in immunity by regulating the expression of autoantigens and negative selection of autoreactive T-cells in the thymus. Mutations in this gene cause the rare autosomal-recessive systemic autoimmune disease termed autoimmune polyendocrinopathy with candidiasis and ectodermal dystrophy (APECED). [provided by RefSeq, Jun 2012]

Uniprot Description

AIRE: Transcriptional regulator that binds to DNA as a dimer or as a tetramer, but not as a monomer. Binds to G-doublets in an A/T-rich environment; the preferred motif is a tandem repeat of 5'-. ATTGGTTA-3' combined with a 5'-TTATTA-3' box. Binds to nucleosomes. Binds to chromatin and interacts selectively with histone H3 that is not methylated at 'Lys-4', not phosphorylated at 'Thr-3' and not methylated at 'Arg-2'. Functions as a sensor of histone H3 modifications that are important for the epigenetic regulation of gene expression. Functions as a transcriptional activator and promotes the expression of otherwise tissue-specific self-antigens in the thymus, which is important for self tolerance and the avoidance of autoimmune reactions. Defects in AIRE are a cause of autoimmune poly- endocrinopathy candidiasis ectodermal dystrophy (APS1). An autosomal recessive disease characterized by the combination of chronic mucocutaneous candidiasis, hypoparathyroidism and Addison disease. Symptoms of mucocutaneous candidiasis manifest first, followed by hypotension or fatigue occurring as a result of Addison disease. APS1 is associated with other autoimmune disorders including diabetes mellitus, vitiligo, alopecia, hepatitis, pernicious anemia and primary hypothyroidism. Most of the mutations alter the nucleus-cytoplasm distribution of AIRE and disturb its association with nuclear dots and cytoplasmic filaments. Most of the mutations also decrease transactivation of the protein. The HSR domain is responsible for the homomultimerization activity of AIRE. All the missense mutations of the HSR and the SAND domains decrease this activity, but those in other domains do not. The AIRE protein is present in soluble high-molecular-weight complexes. Mutations in the HSR domain and deletion of PHD zinc fingers disturb the formation of these complexes. 4 isoforms of the human protein are produced by alternative splicing.

Protein type: Transcription factor

Chromosomal Location of Human Ortholog: 21q22.3

Molecular Function: chromatin binding; histone binding; protein binding; transcription cofactor activity; zinc ion binding

Biological Process: immune response; positive regulation of transcription from RNA polymerase II promoter; positive regulation of transcription, DNA-dependent; regulation of transcription, DNA-dependent

Disease: Autoimmune Polyendocrine Syndrome, Type I, With Or Without Reversible Metaphyseal Dysplasia

Research Articles on AIRE

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Product Notes

The AIRE aire (Catalog #AAA178480) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-AIRE Antibody reacts with Human, Rat No cross reactivity with other proteins. and may cross-react with other species as described in the data sheet. AAA Biotech's AIRE can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). Western Blot: 0.1-0.5mug/ml; Tested Species: Human, Rat Tested Species:In-house tested species with positive results. Other applications have not been tested. Researchers should empirically determine the suitability of the AIRE aire for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "AIRE, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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