Streptavidin allophycocyanin

Manufactured by BioLegend

Streptavidin-allophycocyanin is a fluorescent conjugate that consists of the protein streptavidin coupled to the fluorescent dye allophycocyanin. Streptavidin is a tetrameric protein that binds to the small molecule biotin with high affinity. This conjugate can be used to detect and quantify biotinylated molecules in various applications, such as flow cytometry, immunoassays, and Western blotting.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Allophycocyanin (APC)-conjugated streptavidin Streptavidin-allophycocyanin (SA-APC); Allophycocyanin-labeled streptavidin Streptavidin-allophycocyanin (APC) Allophycocyanin-conjugated streptavidin

4 protocols using streptavidin allophycocyanin

Multiparametric Flow Cytometry Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following mAbs were used for staining and flow cytometric measurements: CD3-VioGreen (BW264/56), CD4-allophycocyanin-Vio770 (VIT4), CD8-PE-Vio770 (BW135/80), CD14-peridinin chlorophyll protein (PerCP) (TÜK4), CD20-PerCP (LT20), CD27-FITC (M-T271), CD28-FITC (15E8), CD45RA-VioBlue (T6D11), CD45RO-FITC (UCHL1), CD62L-FITC (145/15), CD107a-PE (1D4B), CD127-FITC (MB15-18C9), CD137-allophycocyanin, CD137-PE (4B4-1), CD154-VioBlue (5C8), CD178-PE (NOK-1), CD197 (CCR7)-allophycocyanin, CD197 (CCR7)-PE (150503), CD279-allophycocyanin (PD1.3.1.3), anti-IL-2-PE-Vio770 (N7.48A), anti-IL-4-PE (7A3-3), anti-IL-17-FITC (CZ8-23G1), anti-IFN-γ-FITC, anti-IFN-γ-PE-Vio770 (45-15), anti-TNF-α-PE, and anti-TNF-α-VioBlue (cA2) (all Miltenyi Biotec).
Soluble biotinylated pHLA-A*0201 molecules loaded with WT1₃₇ (VLDFAPPGA), WT1₁₂₆ (RMFPNAPYL), WT1₁₈₇ (SLGEQQYSV), WT1₂₃₅ (CMTWNQMNL), or cytomegalovirus (CMV) pp65₄₉₅ (NLVPMVATV) were produced as described previously.14 (link) Tetramerization was achieved by binding to streptavidin-PE or streptavidin-allophycocyanin (both BioLegend, San Diego, CA). In each case, 2·5 × 10⁶ cells were stained with 1 µg/ml tetramer for 45 min at 4°C.
Data were acquired using a MACSQuant flow cytometer and analysed with MACSQuantify software (both Miltenyi Biotec).

Schmied S., Gostick E., Price D.A., Abken H., Assenmacher M, & Richter A. (2015). Analysis of the functional WT1-specific T-cell repertoire in healthy donors reveals a discrepancy between CD4+ and CD8+ memory formation. Immunology, 145(4), 558-569.

+ Open protocol

+ Expand

Multi-parameter Flow Cytometry of Immune Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Spleen cells were first incubated with anti-murine FcgRII/III mAb (2.4G2) for 10 min and then stained with saturating concentrations of Alexa Fluor 488–conjugated, allophycocyanin-conjugated, biotin-conjugated, PEconjugated, FITC-conjugated, PE/Cy5-conjugated, or PE/Cy7-conjugated mAbs against IL-17, IL-4, IFN-γ (BD Biosciences), CD4, glucocorticoidinduced TNFR (BioLegend, San Diego, CA), Foxp3, CTLA4, RORδ, GMCSF, and program death-1 (eBioscience, San Diego, CA). Biotinylated primary mAbs were detected using either streptavidin-allophycocyanin (BioLegend), streptavidin-FITC, streptavidin-PE, or streptavidin-PE/Cy5 (BD Biosciences, San Jose, CA). Primary rabbit anti-mouse RGC-32 Ab (Sigma-Aldrich, St. Louis, MO) was detected using secondary FITC-labeled goat anti-rabbit Ab (Santa Cruz Biotechnology, Dallas, TX). For intracellular cytokine staining, cells were stimulated with 50 ng/ml PMA (Sigma-Aldrich) and 1 µg/ml ionomycin (Sigma-Aldrich) for 4 h, and GolgiPlug was added for the last 2 h (BD Biosciences); cells were then stained for CD4 and intracellular cytokines as previously described (22 (link)). Multicolor flow cytometric analyses were performed using an Accuri C6 and LSR II flow cytometer (BDBiosciences).

Rus V., Nguyen V., Tatomir A., Lees J.R., Mekala A.P., Boodhoo D., Tegla C.A., Luzina I.G., Antony P.A., Cudrici C.D., Badea T.C, & Rus H.G. (2017). RGC-32 Promotes Th17 Cell Differentiation and Enhances Experimental Autoimmune Encephalomyelitis. Journal of immunology (Baltimore, Md. : 1950), 198(10), 3869-3877.

+ Open protocol

+ Expand

Immune Cell Profiling of Pancreatic Islets

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse islet cells were stained with anti-CD45 and biotinylated anti-H2Kd (BD Pharmingen) followed by streptavidin-allophycocyanin (BioLegend). β‐cells were identified by autofluorescence (23 (link)). Dead cells were excluded using propidium iodide. The immune cells infiltrating islets and spleens were stained with anti-CD45, anti-CD3, anti-CD4, anti-CD8a, anti-CD44, anti-CD62L, anti-KLRG-1, anti-CD11b, anti-Ly6G (all BioLegend), anti-BrdU, anti-pSTAT3, and anti-pSTAT5 (all BD Bioscience). Data were collected on the FACS Fortessa cell analyzer (BD Bioscience, San Jose, CA, USA) and were analyzed using FlowJo Software version 10 (TreeStar, Inc, Ashland, OR, USA).

Ge T., Jhala G., Fynch S., Akazawa S., Litwak S., Pappas E.G., Catterall T., Vakil I., Long A.J., Olson L.M., Krishnamurthy B., Kay T.W, & Thomas H.E. (2020). The JAK1 Selective Inhibitor ABT 317 Blocks Signaling Through Interferon-γ and Common γ Chain Cytokine Receptors to Reverse Autoimmune Diabetes in NOD Mice. Frontiers in Immunology, 11, 588543.

+ Open protocol

+ Expand

Multimerized Antigen Probes for B Cell Detection

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antigen-specific B cells were detected using biotinylated proteins in combination with different streptavidin-fluorophore conjugates^{42 (link)}. Biotinylated proteins were multimerized with fluorescently labeled streptavidin for 1 h at 4 °C. Full-length spike protein (R&D Systems) was mixed with streptavidin-Brilliant Violet 421 (BioLegend) at a 10:1 mass ratio (for example, 200 ng spike with 20 ng streptavidin; approximately 4:1 molar ratio). Spike RBD (R&D Systems) was mixed with streptavidin allophycocyanin (BioLegend) at a 2:1 mass ratio (for example, 25 ng RBD with 12.5 ng streptavidin; approximately 4:1 molar ratio). Biotinylated influenza hemagglutinin pools (A/Brisbane/02/2018/H1N1, B/Colorado/06/2017; Immune Technology) were mixed with streptavidin-phycoerythrin (BioLegend) at a 6.25:1 mass ratio (for example, 100 ng hemagglutinin pool with 16 ng streptavidin; approximately 6:1 molar ratio). Streptavidin-Brilliant Violet 711 (BD Biosciences) was used as a decoy probe without biotinylated protein to gate out cells that nonspecifically bind streptavidin. Antigen probes for spike, RBD and hemagglutinin were prepared individually and mixed together after multimerization with 5 µM of free D-biotin (Avidity LLC) to minimize potential cross-reactivity between probes.

Apostolidis S.A., Kakara M., Painter M.M., Goel R.R., Mathew D., Lenzi K., Rezk A., Patterson K.R., Espinoza D.A., Kadri J.C., Markowitz D.M., E. Markowitz C., Mexhitaj I., Jacobs D., Babb A., Betts M.R., Prak E.T., Weiskopf D., Grifoni A., Lundgreen K.A., Gouma S., Sette A., Bates P., Hensley S.E., Greenplate A.R., Wherry E.J., Li R, & Bar-Or A. (2021). Cellular and humoral immune responses following SARS-CoV-2 mRNA vaccination in patients with multiple sclerosis on anti-CD20 therapy. Nature Medicine, 27(11), 1990-2001.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!