Flow cytometry Anti-bodies Human - CD11b

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).

3 Matching solutions
Start a discussion

Start discussion

No discussions found

Start your discussion

Share your thoughts or question with experts in your field

Start a discussion

Found 3 matching solutions for this experiment

CD11b Antibody, anti-human/mouse, FITC

Miltenyibiotec

Upstream tips	Protocol tips	Downstream tips
	An aliquot (50 μL) of each was mixed with 50 μL of ice-cold PBS containing FITC-conjugated anti-human CD11b antibody (4 μL, undiluted) and incubated (4°C, 30 min, in darkness). After adding 2 mL of FACS lysing solution (BD Biosciences, San Jose, CA), incubation (room temp, 10 min), tubes were centrifuged (300 x g, room temp, 5 min). The cell pellets were re-suspended in 200μL of PBS and stored in 4 °C in the dark until evaluation by flow cytometry.

Protocol tips
An aliquot (50 μL) of each was mixed with 50 μL of ice-cold PBS containing FITC-conjugated anti-human CD11b antibody (4 μL, undiluted) and incubated (4°C, 30 min, in darkness). After adding 2 mL of FACS lysing solution (BD Biosciences, San Jose, CA), incubation (room temp, 10 min), tubes were centrifuged (300 x g, room temp, 5 min). The cell pellets were re-suspended in 200μL of PBS and stored in 4 °C in the dark until evaluation by flow cytometry.

Protocol tips

An aliquot (50 μL) of each was mixed with 50 μL of ice-cold PBS containing FITC-conjugated anti-human CD11b antibody (4 μL, undiluted) and incubated (4°C, 30 min, in darkness). After adding 2 mL of FACS lysing solution (BD Biosciences, San Jose, CA), incubation (room temp, 10 min), tubes were centrifuged (300 x g, room temp, 5 min). The cell pellets were re-suspended in 200μL of PBS and stored in 4 °C in the dark until evaluation by flow cytometry.

Manufacturer protocol Publication protocol 1 Relevant paper

CD11b Antibody, anti-human, PE, REAfinity™

Miltenyibiotec

Upstream tips	Protocol tips	Downstream tips
	An aliquot (50 μL) of each was mixed with 50 μL of ice-cold PBS containing FITC-conjugated anti-human CD11b antibody (4 μL, undiluted) and incubated (4°C, 30 min, in darkness). After adding 2 mL of FACS lysing solution (BD Biosciences, San Jose, CA), incubation (room temp, 10 min), tubes were centrifuged (300 x g, room temp, 5 min). The cell pellets were re-suspended in 200μL of PBS and stored in 4 °C in the dark until evaluation by flow cytometry.

Protocol tips
An aliquot (50 μL) of each was mixed with 50 μL of ice-cold PBS containing FITC-conjugated anti-human CD11b antibody (4 μL, undiluted) and incubated (4°C, 30 min, in darkness). After adding 2 mL of FACS lysing solution (BD Biosciences, San Jose, CA), incubation (room temp, 10 min), tubes were centrifuged (300 x g, room temp, 5 min). The cell pellets were re-suspended in 200μL of PBS and stored in 4 °C in the dark until evaluation by flow cytometry.

Protocol tips

Manufacturer protocol Publication protocol 1 Relevant paper

CD11b Monoclonal Antibody (ICRF44), APC, eBioscience™

eBioscience

Upstream tips	Protocol tips	Downstream tips
	cells were incubated on ice for 30min with appropriate antibodies or isotype controls, washed twice with FACS buffer (PBS containing 0.1% NaN3 and 5% fetal bovine serum), and resuspended in 0.5 ml 1% formalin/PBS.	All analyses were performed on a FACS calibur system (Becton Dickinson Immunocytometry Systems).

Protocol tips
cells were incubated on ice for 30min with appropriate antibodies or isotype controls, washed twice with FACS buffer (PBS containing 0.1% NaN3 and 5% fetal bovine serum), and resuspended in 0.5 ml 1% formalin/PBS.
Downstream tips
All analyses were performed on a FACS calibur system (Becton Dickinson Immunocytometry Systems).

Manufacturer protocol Publication protocol 1 Relevant paper

Can't find the product you've used to perform this experiment? It would be great if you can help us by Adding a product!

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

Outsource experiment