SKU: 94069533944

AO/EB Double Staining Kit

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Description

AO/EB Double Staining KitProduct Specification Usage Self prepared material: Fluorescence microscope; PBS; Cell counting board; Glass slide; Cover glass slides 1. Preparation of staining working solution Before the formal experiment, take acridine orange staining solution (reagent A), ethidium bromide staining solution (reagent B), and dilution buffer solution (reagent C), and dilute them to the required AO EB staining working solution in the ratio of A: B: C=1: 1: 8 2. Cell

Product Specification

Usage

Self prepared material:
Fluorescence microscope; PBS; Cell counting board; Glass slide; Cover glass slides

1. Preparation of staining working solution
Before the formal experiment, take acridine orange staining solution (reagent A), ethidium bromide staining solution (reagent B), and dilution buffer solution (reagent C), and dilute them to the required AO/EB staining working solution in the ratio of A: B: C=1:1:8
2. Cell preparation
Adherent cells: Digest according to conventional methods. Centrifuge at 1000 rpm for 5 minutes, collect cell precipitates, and wash once with PBS6pieces/ml
3. Staining
Every 25μ Add 2&mu to the cell suspension; Freshly prepared AO/EB staining working solution, gently mix evenly. Incubate at room temperature for 5-10 minutes
4. Observation
Take a clean slide and add 5-10&mu dropwise; Cover the stained cell suspension with a cover glass. Observe and count using a fluorescent microscope's luciferin filter and a 60x magnification objective. This measurement should be repeated at least three times, with at least 100 total cells observed each time

Description

  Acridine orange (AO) is a heterochromatic nucleic acid binding dye with cell membrane permeability. It binds to DNA or RNA by insertion or electrostatic attraction. When combined with double stranded DNA (dsDNA), it emits green fluorescence (ex/em=502/525nm), and when combined with single stranded DNA (ssDNA) or RNA, it emits red fluorescence (ex/em=460/650nm), with a small amount of binding showing orange red fluorescence. Ethidium bromide (EB) is a polycyclic aromatic hydrocarbon fluorescent small molecule and nucleic acid insertion agent, which is chimeric into the base pairs of double stranded DNA or RNA. It has no base specificity and shows red fluorescence ((ex/em=518/605nm). EB is not cell membrane permeable
  Acridine orange (AO) and ethidium bromide (EB) are often used in combination to observe the changes of nucleus and the formation of apoptotic bodies, which are the characteristics of apoptosis. The combination of the two can distinguish living cells, apoptotic cells and necrotic cells. The working principle is that acridine orange can stain both live and dead cells, while EB can only stain cells that have lost membrane integrity. The living cells are uniformly green. Early apoptotic cells are green, and bright green dots are seen in the nuclear energy of cells due to chromosome condensation and nuclear breakage. Late apoptotic cells will also be stained with EB and thus become orange red, but compared with necrotic cells, late apoptotic cells will have concentrated and often broken nuclei. Necrotic cells are also stained orange red, but will have a nuclear morphology similar to that of living cells, without condensed chromosomes.

Product Components:

Component Name Specification Save
Component A Ao stain solution acridine orange dye solution 200ul 2-8 ℃, protected from light
Component B

EB stain solution ethidium bromide dye solution

200ul 2-8 ℃, protected from light
Component C

Dilution buffer dilution buffer

50ml 2-8 ℃, protected from light
General Notes

1. It is also commonly used to double stain cells with H O echst 33342, which can produce blue fluorescence after binding to dn a through the living cell membrane, and propidiu m iodide, which can only produce red fluorescence after binding to dn a through dead cells
2. If there is a low-temperature centrifuge for centrifugation, the effect is better
3. During operation, pay attention to reducing the exposure time of reagent (a) and reagent (b) to strong light
4. EB solution has certain toxicity, please be careful
5. For your safety and health, please wear lab clothes and disposable gloves

Storage Temp. 2-8 ℃ protected from light, 6months
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SKU: 94069533944

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Erik
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★★★★★ 5
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Color: Large Set of 4
Colors are bright and my dog loves the squeaky! Good size no worries of choking.
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Reviewed in the United States on September 10, 2025
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Mary Freeman
Natrona Heights, US
★★★★★ 5
amazing
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good buy worked well
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Reviewed in the United States on April 13, 2026
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Steve
Boise, US
★★★★★ 4
3.5, Not 3.0
Color: Medium Set of 6
These balls were almost exactly what I needed. I was replacing my boy’s favorite ball, which is the 3” squeaky, spikey ball that fits in the chuck it launcher. Unfortunately I saw the shiny object next to it and because there were SIX balls and a dollar less, I failed to see that these are 3.5” balls instead of 3”…. Great balls but no fitty in the slingy. Buyer beware. No chuck it no problem. Save a buck.
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Reviewed in the United States on March 9, 2026
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HollisHoHo
Dallas, US
★★★★★ 5
MY CHIHUAHUA LOVES THESE BALLS!!
Color: Samll Set of 8
My little chihuahua, BamBam, is addicted to balls, especially little balls that squeak! However, any balls that have material on them, BamBam will tear them apart, swallow the material and then get an upset tummy! These balls saved the day! BamBam LOVES them & she cannot tear them apart! They hold up so well! No matter how much BamBam plays with them, they keep squeaking away!
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Reviewed in the United States on July 15, 2025
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Laurens springerslng
Fort Morgan, US
★★★★★ 5
Great thick squeaky ball
Color: Large Set of 2
These are good thick rubbery squeaky balls. My boy loves them! He's 75lbs. Strong chewer. I hav hard time squeezing to squeak, but he doesn't... He loves it!!!
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Reviewed in the United States on March 24, 2026

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