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Description
Calcein-AM/PI Double Stain KitProduct Specification Usage 1. Preparation of working fluid: 1. Preparation of 1 Assay Buffer Take out 10 Assay Buffer from low temperature refrigerator, take out an appropriate amount according to the aseptic conditions of single dosage, and dilute 10 times with deionized water (dH2O) to obtain 1 Assay Buffer. 2. Preparation of 1 dyeing working solution (1) First, return the cryopreserved Calcein AM solution (2mM) and PI solution (1. 5 mM) to room
Product Specification
| Usage |
1. Preparation of working fluid: 1. Preparation of 1 × Assay Buffer Take out 10 × Assay Buffer from low-temperature refrigerator, take out an appropriate amount according to the aseptic conditions of single dosage, and dilute 10 times with deionized water (dH2O) to obtain 1 × Assay Buffer. 2. Preparation of 1 × dyeing working solution (1) First, return the cryopreserved Calcein-AM solution (2mM) and PI solution (1.5 mM) to room temperature for 20-30min. Note: The mother liquor can be dispensed for the first use to reduce the number of repeated freezing and thawing. (2) Add 5μl of Calcein-AM solution (2mM) and 15μl of PI solution (1.5 mM) to 5ml of 1 × Assay Buffer and mix well. At this time, the working solution concentration of Calcein-AM was 2 μM, and the working solution concentration of PI was 4.5 μM. Because the optimal staining conditions of different cell lines are different, it is recommended to do gradient experiments for the initial experiment to determine the optimal concentrations of Calcein-AM and PI. The principle of gradient screening is to use the lowest probe concentration to obtain the best fluorescence results. Note: Due to the poor stability of Calcein-AM, this dyeing solution must be prepared immediately and used up the same day. 2. Dyeing steps: 1. For adherent cells, first digest the cells with a cell spatula or trypsin-EDTA, and then collect the cells by centrifugation (1000rpm, 3min). For suspended cells, cells were collected by direct centrifugation (1000 rpm, 3 min). 2. Remove the supernatant and fully wash the cells 2 to 3 times with 1 × Assay Buffer to fully remove the residual esterase activity. 3. Prepare the cell suspension with 1 × Assay Buffer to have a density of 1 × 105 ~ 1 × 106 cells/ml. 4. Take 100μl of staining working solution and add it to 200μl of cell suspension, mix well, and incubate at 37 °C for 15min. Note: Incubation time can be extended to 30min if desired. 5. Use a 490 ± 10 nm excitation filter under a fluorescence microscope to simultaneously detect living cells (yellow-green fluorescence) and dead cells (red fluorescence). In addition, only dead cells were observed using a 545 nm emission filter. Detection can also be performed directly under a fluorescent microplate reader using a suitable filter. Note: The following methods can be used to optimize the optimum working concentrations for the two fluorescent dyes. (1) incubating the cells with 0.1% saponin or 0.1-0.5% digoxin for 10 minutes, or incubating the cells with 70% ethanol for 30 minutes, thereby preparing dead cells; (2) Dead cells were stained with 0.1-10 μM PI solution to obtain the optimal working concentration for staining only the nucleus, but not the cytoplasm. (3) Dead cell staining was performed with 0.1-10 μM Calcein-AM to obtain an optimal working concentration that would not stain the cytoplasm. Live cells are then stained at this concentration to see if the live cells can be stained |
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| Synonym | Calcein-AM/PI | ||||||||||||||||
| Description | Calcein-AM is a cell staining reagent that fluorescently labels living cells and fluoresces green (Ex = 490nm, Em = 515nm). Because it introduces acetylmethoxymethyl ester (AM) group on the basis of traditional Calcein, it increases hydrophobicity and makes it easily penetrate living cell membranes. Once in the cell, Calcein-AM (which itself does not fluoresce) is cleaved by esterases in the cell to form a membrane-impermeable polar molecule Calcein, which is retained in the cell and emits strong green fluorescence. Compared with other similar reagents (such as BCECF-AM and CFDA), Calcein, AM is the most suitable fluorescent probe for staining viable cells due to its extremely low cytotoxicity, and does not inhibit any cellular functions, such as proliferation and lymphocyte chemotaxis. Because dead cells lack esterase, Calcein-AM is only used for cell viability testing and short-term labeling of living cells. Therefore, Calcein-AM is often used in combination with dead cell fluorescent probes such as propidium iodide (PI) to perform fluorescent double staining of living cells and dead cells at the same time. Propidium iodide (PI) cannot pass through the cell membrane of living cells, but can only pass through the disordered area of the dead cell membrane to reach the nucleus, and embedded into the DNA double helix of the cell to produce red fluorescence (Ex = 535 nm, Em = 617 nm), so PI only stains dead cells. Since both Calcein and PI-DNA can be excited at 490 nm, live and dead cells can be observed simultaneously with fluorescence microscope. With 545 nm excitation, only dead cells can be observed. The working principle of this kit lies in the double dyes of Calcein-AM and PI to perform double staining labeling of living cells and dead cells, so as to analyze the level of living cells and dead cells. According to our optimized experimental system, a single staining of 200μl cell suspension can perform 500 tests. Product Components:
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| General Notes | 1. Since Calcein-AM is very sensitive to humidity, if the required amount of Calcein-AM solution is taken every time, the lid must be tightly sealed. It is recommended to package and seal storage according to the single dosage. The Calcein-AM working fluid must be prepared for ready use. 2. Propidium iodide (PI) has certain carcinogenicity, so we must pay attention to protection during operation. If it comes into contact with the skin, it needs to be washed with tap water immediately. 3. For your safety and health, please wear a lab coat and disposable gloves. |
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| Instructions | 1. Preparation of working solution: 1, 1× The preparation of Assay Buffer (reaction buffer) Remove from the low temperature refrigerator for 10 times; Assay Buffer, according to the single dosage of sterile conditions to take out the appropriate amount, with deionized water (dH2O) to do 10 times dilution to get 1times; Assay Buffer. 2, 1× Dyeing the configuration of the working liquid (1) the Calcein - AM solution to be kept at low temperature (2 mM) and PI solution (1.5 mM) back to the room temperature 20-30 min. Note: The mother liquor can be divided for the first use to reduce the number of repeated freezing-thawing. (2) apply 5 & mu; l Calcein-AM solution (2mM) and 15μ l PI solution (1.5mM) added to 5ml 1× Assay Buffer and mix well. At this time, the working solution concentration of Calcein-AM obtained is 2μ M, PI working liquid concentration is 4.5 & mu; M. As the optimal staining conditions differ for different cell lines, gradient experiments are recommended for initial experiments to determine the optimal concentration of Calcein-AM and PI. The principle of gradient screening is to use the lowest probe concentration to obtain the best fluorescence results. Note: Due to the poor stability of Calcein-AM, this staining solution must be used on the go and used up the same day. II. Staining steps: 1. For adherent cells, first digest the cells with a cell scraper or pancrease-EDTA, and then collect the cells by centrifugation (1000rpm, 3min). For suspended cells, cells were collected by centrifugation directly (1000 rpm, 3min). 2, remove the supernatant and use 1× The cells were thoroughly washed 2 to 3 times with Assay Buffer to fully remove residual esterase activity. 3, with 1× The cell suspension was prepared with Assay Buffer to a density of 1× 105~1× 106 cells /ml. 4, take 100μ l Add 200&mu to staining working solution; l cell suspension was mixed and incubated at 37 ° C for 15min. Note: The incubation time can be extended to 30min if desired. 5, use 490± under fluorescence microscope; 10 nm excitation filter was used to detect both live cells (yellow-green fluorescence) and dead cells (red fluorescence). In addition, only dead cells could be observed using the 545nm emission filter. It can also be detected directly under a fluorescent microplate reader using a suitable filter. Note: The following methods can be used to optimize the optimal working concentration of the two fluorescent dyes. (1) Dead cells were prepared by incubating cells with 0.1% saponin or 0.1-0.5% digoxin for 10min or 70% ethanol for 30min; (2) use 0.1-10μ M's PI solution was used for dead cell staining to obtain an optimal working concentration that stained only the nucleus, but not the cytoplasm. (3) with 0.1-10μ M of Calcein-AM was used for dead cell staining to obtain an optimal working concentration that would not stain the cytoplasm. This concentration was then used for live-cell staining to see if viable cells could be stained |
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| Storage Temp. | Stored dry at-20 ℃ in the dark from light, the shelf life is 12 months. |
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4.6 ★★★★★
Based on 1315 reviews
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Product Reviews
★★★★★ 5
smells luxurious
smells so luxurious. i am on my third bottle and just purchased the fragance as well as the bar soap. thats how much i love the fragance. i wills say, its really intense smell so dont recommend if you are sensitive to strong/lingering smells. leaves my skin super soft. great value for the money, honestly suprised that is not more expensive. its advertised for men but i use it too and love. definetly good for women too who like intense woody/smoky scents.
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Reviewed in the United States on December 19, 2025
★★★★★ 4
Great Value
You are getting a lot for the money when you get this body wash. It smells great and is an overall wonderful product.
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Reviewed in the United States on March 7, 2026
★★★★★ 5
Smells so good
Love this stuff! Very concentrated. A little goes a long way. Smells amazing and leaves your skin soft and smooth.
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Reviewed in the United States on April 9, 2026
★★★★★ 5
A major upgrade from standard drugstore brands—sophisticated scent & better lather.
I have been a long-time user of standard body washes (think Old Spice, Axe, or Dove Men), but I kept seeing hype about Cremo and decided to see if it was worth the slightly higher price tag. After using the 32oz Spice & Black Vanilla for a week, I can confidently say this is a massive upgrade for your shower routine.
Here is how it stacks up against the typical grocery store brands:
1. The Scent is on another level:
Most cheaper washes smell like generic "Sport" or "Ocean" cologne. This scent is much more complex and earthy.
Top Notes: It starts with a sharp, spicy kick (that's the cardamom).
Finish: It settles into a smooth, warm vanilla and tobacco scent. It smells more like a high-end department store fragrance than a soap.
Staying Power: The scent actually lingers. If you shower in the morning, you will still catch subtle hints of the warm spices for a few hours, which pairs really well with woody colognes.
2. Performance & Feel:
The consistency is thicker than what I'm used to. You only need one solid pump to get a rich lather with a loofah. Crucially, it rinses clean without leaving that "squeaky" residue that dries out your skin, but it also doesn't leave a slick oily film like some ultra-moisturizing washes do. It hits a perfect balance.
3. Value:
The 32oz bottle is substantial. Because the gel is concentrated, I expect this bottle to last 3-4 months easily, making the cost per ounce very reasonable compared to buying smaller bottles of cheaper stuff every few weeks.
Pros:
Sophisticated, "grown-up" scent profile.
High-quality pump (sturdy, doesn't clog).
Non-drying formula is great for colder months.
Cons:
Potency: The scent is strong. If you are sensitive to fragrance or prefer unscented products, this might be overwhelming. The bathroom will smell like it for a good 20 minutes after you're done (which I enjoy, but keep it in mind).
Verdict:
This bridges the gap between basic hygiene and luxury grooming. If you are tired of smelling like "Blue Sport" and want something warmer and more refined for the fall/winter, this is absolutely worth the switch.
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Reviewed in the United States on November 26, 2025
★★★★★ 5
It smells as good as my cologne.
I never go out without a shot of Guerlain L'Homme Idéal L'Intense cologne. Guerlain does not offer a layered line of products with similar scents. I wanted a deodorant and body wash to match. None of the Old Spice scents came close. Axe Oud Wood & Dark Vanilla antiperspirant has more vanilla and is a bit subdued but works.
Finding a body wash was harder. Cremo Spice & Black Vanilla Body Wash sounded promising, but I was apprehensive about its scent including tobacco. I did not want to smell like an ashtray. But when I opened the bottle, I was met with a surprising scent from the past. I immediately recognized it as premium Danish pipe tobacco. Most people do not recognize this aromatic scent as coming from tobacco at all. In fact, it smells more like fresh cured tobacco itself, not its smoke. This adds a very pleasant and masculine smell to the body wash on top of the spice, wood, and vanilla scent I was searching for.
The scent only lasts a few hours after you rinse off. One push on the dispenser puts enough soap on a shower loofah to wash your whole body. I do not rinse off my loofah; I squeeze the soap out of it to wash my face, then I place it back in its tray next to the bottle. The scent of the remaining soap acts like incense until my next shower.
I was also pleasantly surprised to find that this body soap has a rich lather and cleans as well as advertised. I had forgotten the reason I bought the body soap until I used it.
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Reviewed in the United States on February 27, 2026
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