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Home > ÀüÁ¦Ç°º¸±â > Çü±¤ ´Ü¹éÁú ¡¤ Reporter > Çü±¤ ´Ü¹éÁú °ü·ÃÁ¦Ç° > Flow Cytometer Calibration Beads for AcGFP/EGFP and mCherry

Flow Cytometer Calibration Beads for AcGFP/EGFP and mCherry

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Clontech
632594
AcGFP Flow Cytometer Calibration Beads
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20 Assays
663,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32670
Clontech
632595
mCherry Flow Cytometer Calibration Beads
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20 Assays
663,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x32670

Features
Mixture of six discrete bead populations having distinct fluorescent intensities:
  • The lowest intensity represents the autoflorescence signal of cells not expressing the fluorescent protein (AcGFP1 or mCherry)
  • The remaining five peaks are evenly distributed over the remaining scale of the green or red fluorescence detection channel

Use these beads to calibrate your flow cytometer prior to analyzing cells that express the AcGFP1 or mCherry fluorescent proteins. The AcGFP Flow Cytometer Calibration Beads allow for easy calibration of any flow cytometer with a laser line that excites the green fluorescent proteins AcGFP1 (Aequorea coerulescens GFP) and EGFP. [The AcGFP Flow Cytometer Calibration Beads work for both AcGFP1 and EGFP because their excitation/emission spectra and brightness are almost identical.] The mCherry Flow Cytometer Calibration Beads allow for easy calibration of any flow cytometer with a laser line that excites the red fluorescent protein, mCherry.

Each bead suspension contains six distinct populations of beads that vary in the number of attached AcGFP1 or mCherry molecules, which gives each population a distinct fluorescence intensity. The Molecular Equivalent of Soluble Fluorophore (MESF) value for each peak was determined by correlating the fluorescence intensity of each respective bead population with the amount of soluble AcGFP1 or mCherry yielding the same fluorescence intensity.

Converting fluorescence intensity readings to MESF makes it possible to estimate and compare relative expression levels between cells in the same cell population or in independent samples, and even to compare data from different instruments and experiments. Using the Flow Cytometer Calibration Beads, you can also determine the linearity and stability of your flow cytometer¡¯s readouts. The lowest intensity peak represents the autofluorescence signal of cells not expressing the green or red fluorescent protein; this allows you to measure the fluorescence detection threshold and the background noise level of your flow cytometer. The five remaining peaks are evenly distributed over the remaining scale of the fluorescence detection channel.



±×¸² 1. Flow cytometer analysis of AcGFP Flow Cytometer Calibration Beads. 20 ¥ìl of the AcGFP Flow Cytometer Calibration Bead suspension was thorougly resuspended in 1 ml of 1x Flow Cytometer Calibration Beads Dilution Buffer. The bead suspension was then analyzed on a FACS Calibur (BD) flow cytometer using the 488 nm laser line and detecting green fluorescence in the FL-1 channel. 60,000 events were analyzed. The obtained graph shows that the bead suspension contains 6 well-distinguished populations with different fluorescent intensities.



±×¸² 2. Flow cytometer analysis of mCherry Flow Cytometer Calibration Beads. 20 ¥ìl of the mCherry Flow Cytometer Calibration Bead Suspension was thoroughly resuspended in 1 ml of 1x Flow Cytometer Calibration Beads Dilution Buffer. The bead suspension was then analyzed on a FACS Diva (BD) flow cytometer using the 561 nm laser excitation line and detecting in the red channel. 10,000 events were analyzed. This graph shows that the bead suspension contains 6 well-distinguished populations with different fluorescent intensities.
Applications
  • Calibrate your flow cytometer prior to analyzing cells expressing AcGFP1 or mCherry fluorescent protein
  • Since the spectral properties and brightness of AcGFP1 and EGFP are very similar, these beads may be used to calibrate flow cytometers prior to using EGFP expressing cells

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