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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S. Berg
Boise, US
★★★★★ 5
Good balls
Color: S6-glow
My dog is a chewer, and these have held up for close to a year actually a pretty good product
WAS THIS REVIEW HELPFUL?YesReportShare
Reviewed in the United States on May 27, 2026
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nanny america
Charlottesville, US
★★★★★ 3
Disappointed
Color: S6-muti-color
These were great for about 20 minutes then they stopped squeaking...we have 2 left that I have saved but the other 4 do not squeak anymore...disappointed as my dog loves to squeak things...would not buy these again...My dog is only an 8lb dog too...but he can still play with the balls without the squeak...
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Reviewed in the United States on April 26, 2026
J
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joymom
Louisville, US
★★★★★ 5
Great Value for a ball my dog loves!
Color: S6-muti-color
I wish these ball were in the subscription program! My dog gets sooo excited by the new ball, but her interest in things lasts for a few days. She kills the squeak within 11minutes, but the squeaker stays in place rather than becoming a chocking or swallow hazard, and she still loves to chew it. Next, we must throw the ball approximately 723 times the first day, possibly 496 times on day 2, and just 3- 4 on day 3, at which point she will chase the ball before hollering "it's over here if you need it," while she checks on the chipmunk den. We currently have 17 of these balls in our yard (because we have given several leftovers to a less discriminating doodle next door). These balls hold up really well, get her extremely active for the first couple of days, and are a much cheaper than doggie daycare (after an active morning with a new ball, she is happy to chew on it and sleep much of the day). I always have these on hand!
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Reviewed in the United States on November 22, 2025
M
Verified Purchase
Michelle
Bozeman, US
★★★★★ 4
Pieces chewed off
Color: white & green
My Aussiedoodle loves it. I throw it down the hallway, it bounces crazy, and he chases it. The only complaint is that he can't chew much of the ball part. So now I have to have it put away and brought out for special playtime.
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Reviewed in the United States on April 9, 2026
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DDT
Birmingham, US
★★★★★ 5
Great fun for our dogs!
Style: Original
My dogs can’t get enough of this very noisy ball. We have one outside and one in their playroom. It is very noisy. It is very hard. So, I’d say a room with fine furniture is not the place for it. Ours bang it around pretty good. Our boxer did split one, biting it. But, she’s a serious chewer! Our other dog carries it around sometimes but hasn’t been able to damage it yet. So, I’d say for most chewers it’s going to last a long time. It’s a great toy but in most households I’d say basement or outside. Again, it’s hard and did ding a few baseboards.
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Reviewed in the United States on March 27, 2026

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