Flow cytometry Anti-bodies Mouse - CD138/Syndecan-1

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

PE Rat Anti-Mouse CD138

BD Biosciences

Upstream tips	Protocol tips	Downstream tips
Mice were euthanized by approved methods and then tissues were immediately removed from the mice and placed in FACS buffer on ice.	FACS buffer consists of DPBS with 0.5% Bovine Serum Albumin and 2mM EDTA. Cells were incubated on ice in 100 µl of antibody cocktail for 20 minutes followed by two washes with FACS buffer.	The cell suspension was brought up in 400 µl of FACS buffer then run on a LSR II flow cytometer (BD Biosciences, configuration in Supplementary Table 1).

Upstream tips
Mice were euthanized by approved methods and then tissues were immediately removed from the mice and placed in FACS buffer on ice.
Protocol tips
FACS buffer consists of DPBS with 0.5% Bovine Serum Albumin and 2mM EDTA. Cells were incubated on ice in 100 µl of antibody cocktail for 20 minutes followed by two washes with FACS buffer.
Downstream tips
The cell suspension was brought up in 400 µl of FACS buffer then run on a LSR II flow cytometer (BD Biosciences, configuration in Supplementary Table 1).

Manufacturer protocol Publication protocol 1 Relevant paper

Purified anti-mouse CD138 (Syndecan-1) Antibody

BioLegend

Upstream tips	Protocol tips	Downstream tips
Single cell suspension of spleens and bone marrow (femurs and tibia) were prepared and filtered through a 70 μm cell strainer (BD Falcon). The primary cells were resuspended (107 cells/ml) in PBS containing 0.5%. BSA 0.5%.		After washing twice, cells were re-suspended in PBS/0.5% BSA/2 mM EDTA, and analyzed in an LSRII flow cytometer (BD Biosciences). The resulting data were analyzed using the FlowJo software.

Upstream tips
Single cell suspension of spleens and bone marrow (femurs and tibia) were prepared and filtered through a 70 μm cell strainer (BD Falcon). The primary cells were resuspended (107 cells/ml) in PBS containing 0.5%. BSA 0.5%.
Downstream tips
After washing twice, cells were re-suspended in PBS/0.5% BSA/2 mM EDTA, and analyzed in an LSRII flow cytometer (BD Biosciences). The resulting data were analyzed using the FlowJo software.

Manufacturer protocol Publication protocol 1 Relevant paper

PE anti-mouse CD138 (Syndecan-1) Antibody

BioLegend

Upstream tips	Protocol tips	Downstream tips
	Cells were surface-stained, then fixed/permeabilized (Fix-Perm buffer, eBioscience) before intracellular staining.

Protocol tips
Cells were surface-stained, then fixed/permeabilized (Fix-Perm buffer, eBioscience) before intracellular staining.

Manufacturer protocol Publication protocol 1 Relevant paper

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