Cd49b apc clone dx5

Manufactured by BioLegend

CD49b-APC (clone DX5) is a monoclonal antibody that recognizes the CD49b cell surface antigen. It is conjugated with the fluorescent dye Allophycocyanin (APC).

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

CD49b-APC (6 citations) CD49b (DX5; (5 citations)

2 protocols using cd49b apc clone dx5

Comprehensive Tumor and Immune Cell Analysis

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

SA1/N and B16-SIY tumor analyses were performed 2 days following the second dose of antibody treatment. Tumors were weighed on a microscale on plastic weigh boats and dissociated with a 0.25mg/mL Liberase TL (Sigma-Aldrich), 0.05mg/mL DNaseI (Roche) from bovine pancreas, and RPMI Medium mixture for 20 min at 37To determine ICOS receptor quantity on primary cells, tumors were processed as described above. Peripheral blood was collected by cardiac puncture with 25-gauge needles and tuberculin syringes and dispersed into EDTA coated blood collection tubes. RBCs were lysed in conical tubes with 1mL of ACK lysis buffer for 3min on ice. Blood was centrifuged at 400xg for 5mins and resuspended in 100uL FACS buffer. Tumors and peripheral blood were transferred to round bottom 96-well plates and Fc blocked as described above. Following Fc block, cells were stained with 1ug/mL DyLight-650 (Life Technologies) labelled ICOS antibody 37A10 or isotype control, in addition to the staining cocktail described above. Cells were stained in 100uL of cocktail for 30mins at 4ºC. Cells were then washed twice with FACS buffer and resuspended 100uL in Fix/Perm buffer (Foxp3/Transcription Factor Staining Kit, eBioscience) and kept at 4ºC for 15mins. Cells were washed twice in Perm buffer and resuspended in 100uL of intracellular staining cocktail containing fluorescent labelled anti-FoxP3 (FJK-16s, 1:100, eBioscience) diluted in Perm buffer. Following a 30min incubation at 4ºC, cells were washed twice in FACS buffer and resuspended in 100uL of FACS buffer supplemented with 0.1% paraformaldehyde. Receptor quantitation was performed using Quantum Simply Cellular beads (Bangs Laboratories) according to manufacturer’s protocol. To determine the number of cells per milligram of tumor, a fixed number of CountBrite beads (Life Technologies) were added to samples and analyzed by flow cytometry.
For TIL analysis of MC38 tumors, huCTLA-4 mice were randomized in groups when tumor reached an average size of 153mm³ and treated with a single dose of clinical grade ipilimumab (Yervoy®, Bristol-Meyers Squibb) or human IgG1 isotype control administered intravenously at 10 mg/kg, or a single dose of ICOS antibody or mouse IgG2a isotype control administered intraperitoneally at 0.25 mg.kg. Mice were sacrificed 72 hours later and tumors excised, dissociated, and processed for flow cytometry analysis. The following antibodies were used: CD45-BB515 (clone 30-F11, BD), CD3-BUV395 (clone 145-2C11, BD), CD4-APCeF780 (clone GK1.5, eBiosciences), CD8-PEeF610 (clone 53–6.7, eBiosciences), FoxP3-PE (clone NRRF-30, eBiosciences), CD25-PerCP/Cy5.5 (clone PC61, Biolegend), CD19-BV605 (clone 6D5, Biolegend), CD49b-APC (clone DX5, Biolegend), ICOS-BV421 (clone C398.4A, Biolegend). Viability was assessed by staining with Fixable Viability Dye eF506 (Biolegend).

Hanson A., Elpek K., Duong E., Shallberg L., Fan M., Johnson C., Wallace M., Mabry G.R., Sazinsky S., Pepper L., Shu C.J., Sathyanarayanan S., Zuerndorfer S., Simpson T., Gostissa M., Briskin M., Law D., Michaelson J, & Harvey C.J. (2020). ICOS agonism by JTX-2011 (vopratelimab) requires initial T cell priming and Fc cross-linking for optimal T cell activation and anti-tumor immunity in preclinical models. PLoS ONE, 15(9), e0239595.

+ Open protocol

+ Expand

Comprehensive Immune Cell Profiling

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

CD3-FITC clone 145-2C11, CD4-APC-Cy7 clone RM4-5, CD8-FITC clone 53-6.7, CD45-AF700 clone 104, CD11b-PE-Cy7 clone M1/70, CD11c-BV450 clone N418, CD19-PerCP-Cy5.5 clone 6D5, CD49b-APC clone DX5, F4/80-BV421 clone BM8, IL-17A-PE-Cy7 clone TC11-18H10.1, Ly6G-APC clone 1A8, and TCRγδ-BV421 clone GL3, and were purchased from Biolegend (San Diego, CA). Ly6C-PerCP-Cy5.5 clone AL-21, RORγt-BV650 clone Q31-378, and Siglec F-PE clone E50-2440 were purchased from BD Biosciences (San Jose, CA). Foxp3-PE clone 150D/E4 was purchased from ThermoFisher Scientific (Waltham, MA).

Denny J.E., Powers J.B., Castro H.F., Zhang J., Joshi-Barve S., Campagna S.R, & Schmidt N.W. (2019). Differential Sensitivity to Plasmodium yoelii Infection in C57BL/6 Mice Impacts Gut-Liver Axis Homeostasis. Scientific Reports, 9, 3472.

+ 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!