Thermo Fisher Scientific Nur77 Monoclonal Antibody (12.14), PerCP-eFluor 710, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

0.125 µg/test

View 6 publications 6 publications

Product Specifications

Species Reactivity

Mouse

Published species

Mouse

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

12.14

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

View additional formats

• Unconjugated
• Alexa Fluor 488
• Alexa Fluor 647
• eFluor 450
• PE
• PE-Cyanine7
• Request custom conjugation

Excitation/Emission Max

482/708 nm View spectra

Form

Liquid

Concentration

0.2 mg/mL

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2573815

Product Specific Information

Description: This 12.14 monoclonal antibody reacts with mouse Nur77 (also known as NR4A1, TR3, NGFI-B, or NAK1), an inducible orphan nuclear receptor. Expressed in thymocytes and T cell lines, Nur77 promotes apoptosis and plays a role in thymocyte negative selection. Additionally, Nur77 has been shown to be critical for steroid biosynthesis in Leydig cells as well as for the effects of dopamine. In addition, Nur77 has been shown to interact with FoxP3 in regulatory T cells. However, our results with this antibody do not correlate with this observation.

Applications Reported: This 12.14 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This 12.14 antibody has been tested by intracellular staining followed by flow cytometric analysis of stimulated mouse thymocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to BestProtocols®: Protocol B: One step protocol for (nuclear) intracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.

Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.

Excitation: 488 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

NR4A1 is a member of the steroid-thyroid hormone-retinoid receptor superfamily. NGFI-B-alpha is expressed in brain, adrenal, thyroid, liver, testis, ovary, thymus, muscle, lung and prostate. Its expression is induced in response to various stress stimuli and growth factors. The encoded protein acts as a nuclear transcription factor involved in mediating apoptotic signaling in thymocytes and tumor cells, as well as having a role in signaling in the hypothalamic-pituitary axis. Translocation of the protein from the nucleus to mitochondria induces apoptosis. Three transcript variants encoding two distinct isoforms have been identified for this NR4A1. Variant (3), also known as TRC beta, contains multiple differences compared to variant 1. The resulting isoform b contains a shorter and distinct C-terminus compared to isoform a. RXR has been shown to be a partner for NR4A1.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.