Description
The DF-T1 monoclonal antibody specifically recognizes CD43 which is also known as Leukosialin, Sialophorin, Leukocyte Sialoglycoprotein, Galactoglycoprotein (GALGP), or Ly-48. CD43 is an ~95-135 kDa heavily O-sialylated type I transmembrane glycoprotein that is encoded by SPN which belongs to the cell surface mucin family. CD43 is highly expressed on T lymphocytes, thymocytes, monocytes, granulocytes, bone marrow stem cells, pre-B cells, activated B cells and plasma cells but not on resting peripheral blood B cells, red blood cells, and nonhematopoietic cells. CD43 binds to CD54 (ICAM-1), CD62E (E-selectin), CD169 (Siglec-1), CD328 (Siglec-7), and MHC Class I molecules. It mediates intercellular interactions that regulate leukocyte functions including cellular adhesion and anti-adhesion, lectin binding, co-stimulation of T cell activation and survival, and induction of apoptosis of T cells and hematopoietic progenitors. Enzymatic shedding of CD43 from leukocyte surfaces following their activation by various stimuli may play a negative regulatory role in adaptive immune responses.
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® CompBeads 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 BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. 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.
7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
8. An isotype control should be used at the same concentration as the antibody of interest.
9. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
Specifications

General

SourceHuman B7-H4 amino acids (Phe29-Ala258) (Accession # Q7Z7D3) was expressed in CHO cells.The carboxy terminus contains Fc-6His tag.
Molecular MassThe 475 amino acid recombinant protein has a predicted molecular mass of approximately 53.0 kD. The DTT-reduced and non-reduced protein migrate at approximately 70kD and 140kD respectively by SDS-PAGE. The predicted N-terminal amino acid is Phe.
Purity> 95% by SDS-PAGE gel as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in PBS pH 7.2.
Endotoxin LevelLess than 0.1 EU 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.
ActivityHuman recombinant B7-H4 inhibits the anti CD3 (Cat. No. 317303)-induced proliferation of activated human T cells; the inhibitory effect was measured using Deep Blue Cell Viability™ Kit (Cat. No. 424701). The ED50 = 0.8 - 4 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].
StructureDimer
DistributionAntigen presenting cells (dendritic cells, peritoneal macrophages, and splenic B cells), epithelium of normal tissues breast, pancreas, and kidney, and some tumor cells derived from mammary, ovary, and uterus.
FunctionB7- H4 inhibits T cell activation, proliferation, and cytokine production. B7-H4 is downregulated after B cell activation and induced by L-10 and TGF-β in peritoneal macrophages.
InteractionActivated T cells
BioactivityHuman recombinant B7-H4 inhibits the anti CD3-induced proliferation of activated human T cells.
Biology AreaCostimulatory Molecules, Immunology
Molecular FamilySoluble Receptors
Antigen ReferencesPrasad DV, et al. 2003. Immunity. 18:863. Zang X, et al. 2003. PNAS. 100:10388. Ou D, et al. 2006. Cell Transplant. 15:399. Azuma T, et al. 2009. PLoS Med. 6(10):e1000166. Wei J, et al. 2011. J Exp Med. 208:1683. Radichev IA, et al. 2014. Diabetes. 63:3470. Smith JB, et al. 2016. Mol Ther. 24:1987. Podojil JR, Miller SD. 2017. Immunol Rev. 276:40.
Gene ID79679
UniProtView information about B7-H4 on UniProt.org
Regulatory StatusRUO
Other NamesB7 Homolog 4, B7 Superfamily Member 1 (B7S1), Immune Costimulatory Protein B7-H4 (B7-H4), V-Set Domain Containing T Cell Activation Inhibitor 1 (VCTN1), T Cell Costimulatory Molecule B7x (B7X)