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Invitrogen™ c-Rel Monoclonal Antibody (1RELAH5), eFluor™ 660, eBioscience™
Description
The 1RELAH5 monoclonal antibody reacts with mouse c-Rel. c-Rel is a 65 kDa hematopoietic lineage-restricted member of the NF-kB family of transcription factors. In the thymus, c-Rel is upregulated in regulatory T cells (Treg) when compared to other CD4+ thymocytes. c-Rel is believed to play a critical role in thymic Treg (nTreg) development, but is not necessary for TGF beta-mediated T reg (iTreg) differentiation. C-Rel is also involved in proliferation and survival in mature T cells. Applications Reported: This 1RELAH5 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This 1RELAH5 antibody has been tested by intracellular staining and flow cytometric analysis of mouse thymocytes using the Foxp3 Fixation/Permeabilization Buffers (Product # 00-5521-00) and protocol. This can be used at less than or equal to 0.06 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. 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:...
Specifications
Specifications
| Antigen | c-Rel |
| Applications | Flow Cytometry |
| Classification | Monoclonal |
| Clone | 1RELAH5 |
| Concentration | 0.2 mg/mL |
| Conjugate | eFluor 660 |
| Formulation | PBS with 0.09% sodium azide; pH 7.2 |
| Gene | REL |
| Gene Accession No. | P15307 |
| Gene Alias | BCD541; C-Rel; C-Rel protein; C-Rel proto-oncogene protein; gemin 1; gemin-1; LOW QUALITY PROTEIN: proto-oncogene c-Rel; oncogene REL, avian reticuloendotheliosis; proto-oncogene c-Rel; REL; REL proto-oncogene, NF-kB subunit; reticuloendotheliosis oncogene; SMA; SMA1; SMA2; SMA3; SMA4; SMN; SMN2; SMNT; v-rel avian reticuloendotheliosis viral oncogene homolog; v-rel reticuloendotheliosis viral oncogene homolog |
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Frequently Asked Questions (FAQs)
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.
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).
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.
The eFluor Organic fluorochromes and eVolve QDots can be used with flow staining buffers containing PBS and protein.
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.
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