Flow cytometry Anti-bodies Mouse - TCRbeta

Flow cytometry is an immunophenotyping technique whereby sing cell suspensions are stained for either cell surface markers or intracellular proteins by fluorescently-labelled antibodies and analyzed with a flow cytometer, where fluorescently-labelled molecules are excited by the laser to emit light at varying wavelengths, which is then detected by the instrument. There are several key criteria which are required to be kept in mind while designing a flow experiment- 1. Antibody titration (optimal dilution of antibodies should be calculated in order to avoid over- or under- saturated signals for proper detection of surface and intracellular markers), 2. Precision (3 or more replicates of the sample should be used per experiment), 3. Specificity (proper isotype controls should be included in the experiment), 4. Day-to-day variability (experiments should be repeated 3 or more times to ensure consistency and avoid variability due to flow cytometer settings), 5. Antibody interaction (Fluorescence minus one or FMO should be used, which is the comparison of signals from panel minus one antibody vs. the full panel), and 6. Antibody stability (fluorescently-labelled antibodies should be stored at 4C).

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Found 3 matching solutions for this experiment

TCR beta Monoclonal Antibody (H57-597), FITC, eBioscience™

eBioscience

Upstream tips	Protocol tips	Downstream tips
	7AAD (Biolegend) or Zombie Aqua Fixable Viability Kit (Biolegend) was used to exclude dead cells. For intracellular transcription factor staining, surface-stained cells were fixed and permeabilized using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), according to the manufacturer's protocol	Cells were acquired on the BD FACSCANTO II and LSR (BD Biosciences) and analysis was carried out using FlowJo (Tree Star).

Protocol tips
7AAD (Biolegend) or Zombie Aqua Fixable Viability Kit (Biolegend) was used to exclude dead cells. For intracellular transcription factor staining, surface-stained cells were fixed and permeabilized using the Foxp3/Transcription Factor Staining Buffer Set (eBioscience), according to the manufacturer's protocol
Downstream tips
Cells were acquired on the BD FACSCANTO II and LSR (BD Biosciences) and analysis was carried out using FlowJo (Tree Star).

Manufacturer protocol Publication protocol 1 Relevant paper

Purified Anti-Mouse TCR beta (H57-597)

Tonbo Biosciences

Upstream tips	Protocol tips	Downstream tips
Liver tissues were processed by enzymatic digestion with 2mg/mL type IV collagenase (Worthington, NY). In some experiments, cells were stimulated with 50ng/mL Phorbol 12-Myristate 13-Acetate (PMA; LC Labs, USA) and 1μg/mL Ionomycin (Enzo Life sciences, USA) for 4 hours in the presence of Golgi Stop/Golgi Plug (BD Biosciences).		Cells were analyzed using BD LSRII and FlowJo9.6.4 (TreeStar, Oregon, USA).

Upstream tips
Liver tissues were processed by enzymatic digestion with 2mg/mL type IV collagenase (Worthington, NY). In some experiments, cells were stimulated with 50ng/mL Phorbol 12-Myristate 13-Acetate (PMA; LC Labs, USA) and 1μg/mL Ionomycin (Enzo Life sciences, USA) for 4 hours in the presence of Golgi Stop/Golgi Plug (BD Biosciences).
Downstream tips
Cells were analyzed using BD LSRII and FlowJo9.6.4 (TreeStar, Oregon, USA).

Manufacturer protocol Publication protocol 1 Relevant paper

Purified Hamster Anti-Mouse TCR β Chain

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
	Cell suspensions were stained with antibodies for flow cytometry. For intracellular staining, cells were fixed using Foxp3/Transcription Factor Staining Buffer Set (eBioscience) before staining with intracellular antibodies. For intracellular cytokine secretion assays, cells were incubated for 4 hr with PMA and Ionomycin in the presence of Brefeldin A.

Protocol tips
Cell suspensions were stained with antibodies for flow cytometry. For intracellular staining, cells were fixed using Foxp3/Transcription Factor Staining Buffer Set (eBioscience) before staining with intracellular antibodies. For intracellular cytokine secretion assays, cells were incubated for 4 hr with PMA and Ionomycin in the presence of Brefeldin A.

Manufacturer protocol Publication protocol 1 Relevant paper

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