Description
The LC4 monoclonal antibody specifically recognizes the human variable beta 5.1 (Vβ5.1) domain of the beta subunit for the αβ T cell receptor for antigen (TCR αβ). TCR Vβ5.1 is encoded by TRBV5-1 (T cell receptor beta variable 5-1), one of five functional genes within the TRBV5 subgroup in the T cell receptor beta (TRB) locus. The heterodimeric TCR αβ is composed of two disulfide-linked transmembrane glycoproteins, ie, highly variable TCRα and TCRβ chains. These chains are each comprised of an extracellular N-terminal variable (V) region domain followed by a constant (C) region domain, a transmembrane region, and a short C-terminal cytoplasmic tail. The TCR Vβ repertoire is known to be extensive due to the many different combinations of TCR gene segments (Vβ, Dβ, and Jβ) as well as junctional region diversity. TCR Vβ5.1 is variably expressed on subsets of TCR αβ-positive thymocytes and peripheral CD4+ or CD8+ T cells. In association with the CD3 complex of signaling proteins, the TCR αβ recognizes peptide-major histocompatibility complexes (pMHC) that are displayed on other cells to mediate cellular responses. The LC4 antibody is useful for analyzing the levels of TCR Vβ5.1 expressed by individual cells as well as the numbers or frequencies of TCR Vβ5.1-positive cells within test samples. The LC4 antibody can be used to help characterize the TCR Vβ repertoires of T cell populations during health as well as in response to vaccination, infectious disease, aging, transplantation, autoimmunity or cancer.
BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD CompBead to ensure that BD CompBeads are appropriate for your specific cellular application. For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385). Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
2. Researchers should determine the optimal concentration of this reagent for their individual applications.
3. An isotype control should be used at the same concentration as the antibody of interest.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
9. BD Horizon Brilliant Ultraviolet 496 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
Specifications

General

SourceMouse IFN-α1, amino acids Met-(Cys24-Lys189) (Accession# NM_010502), was expressed in E. coli.
Molecular MassThe 167 amino acid recombinant protein has a predicted molecular mass of approximately 19.2 kD. The protein migrates at approximately 18 kD in DTT-reducing conditions and at 20 kD in non-reducing conditions by SDS-PAGE. The predicted N-terminal amino acid is Met.
Purity>95%, as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in PBS, pH 7.2.
Endotoxin LevelLess than 0.01 ng per µg cytokine as determined by the LAL method.
Concentration10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & HandlingUnopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
ActivityRecombinant Mouse IFN-α1 induces CCL2 (MPC-1) in Raw264.7 cells in a dose dependent manner. The ED50 range for this effect is 0.08 – 0.8 µg/mL
ApplicationBioassay
Application NotesBioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
Product CitationsHu W, et al. 2015. Proc Natl Acad Sci U S A. 112: 13994 - 13999. PubMed Tran NL, et al. 2022. Elife. 11:. PubMed
StructureCytokine
DistributionMonocytes/macrophages, fibroblasts, lymphoblastoid cells, and a variety of cell types following induction by viruses. IFN-α is also induced by stimuli such as nucleic acids, glucocortoid hormones, n-butyrate, and 5-bromodeoxy uridine in different cells.
FunctionIFN-α stimulates cytotoxic T lymphocytes and natural killer cell function. IFN-α has immunomodulatory and antiviral properties.
InteractionReceptors for type I IFNs are ubiquitously present on all cells.
Ligand/ReceptorIFNAR complex has two components: IFNAR1 and IFNAR2.
BioactivityCytopathic effect inhibition assay using EMC virus on A549 cells.
Biology AreaImmunology, Innate Immunity
Molecular FamilyCytokines/Chemokines
Antigen References1. Siren J, et al. 2005. J. Immunol. 174:1932. 2. Bose A and Baral R. 2007. Immunol. Lett. 108:68. 3. Zhao W, et al. 2008. J. Immunol. 180:5483. 4. Sadler AJ and Williams BR. 2008. Nat. Rev. 8:559. 5. Björck P, et al. 2011. J. Immunol. 186:1477. 6. Badiger R, et al. 2012. PLoS One 7:e46779. 7. Aghemo A, et al. 2010. Nat. Rev. Gastroenterol. Hepatol. 7:485. 8. Mukherjee KK, et al. 2012. Indian J. Med. Res. 136:54. 9. Becker-Merok A, et al. 2013. Lupus 22:155.
Gene ID15962
UniProtView information about IFN-alpha1 on UniProt.org
Regulatory StatusRUO
Other NamesIFN-alpha 1, Ifa1, IFN-a1, IFNa1M