Senescence Detection Kit

Histochemical kit

WARNING: This product can expose you to chemicals including Formaldehyde, which is [are] known to the State of California to cause cancer.  For more information go to

K320 is available from Abcam as ab65351.
Catalog #: K320 | abID: ab65351

Product Details

abID ab65351
Cat # +Size K320-250
Size 250 Stainings
Species Reactivity Mammalian
Applications The Senescence Detection Kit is designed to histochemically detect specific senescence marker in distinct pH in cultured cells and tissue sections.
Features & Benefits • Simple procedure; takes around than 30 minutes
• Fast and convenient
• The specific histochemical marker is only present in senescent cells and is not found in presenescent, quiescent or immortal cells.
Kit Components • Fixative Solution (1X)
• X-Gal (150 mg, lyophilized)
• Staining Solution (1X)
• Staining Supplement (100X)
Storage Conditions -20°C
Shipping Conditions Gel Pack
USAGE For Research Use Only! Not For Use in Humans.


Senescence is thought to be a tumor suppressive mechanism and an underlying cause of aging. Senescence represents an arrested state in which the cells remain viable, but not stimulated to divide by serum or passage in culture. Senescent cells display increase of cell size, senescence-associated expression of β-galactosidase (SA-β-Gal) activity, and altered patterns of gene expression. The Senescence detection kit is designed to histochemically detect SA-β-Gal activity in cultured cells and tissue sections, a known characteristic of senescent cells. The SA-β-Gal is present only in senescent cells and is not found in presenescent, quiescent or immortal cells.

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Can this kit be used with tissue section?
The kit has been used for skin sections successfully. Briefly, the tissue was frozen in liquid nitrogen, and mounted in OCT. The thin sections (4 um) were cut, mounted onto glass slides, fixed in 1% formalin in PBS for 1 min at room temp., washed in PBS, immersed overnight in beta-Gal staining solution. Then you can view under bright field at 100-200X magnifications. The staining results can be found in the article below (The reference is also a principal reference describing the senescence marker). Dimri, G.P., et al. (1995) PNAS 92:9363-9367.
Can this kit be used with paraffin section?
This has not been determined as yet.
Does this kit detect transient expression of p53 (3-5 days) or longer termexpression?
The Senescence Detection Kit (K320-250) will detect senescent cells. If the p53 expressing cells become senescent, then the kit should detect it. It does not matter what causes senescence, but as long as cells become senescent, the kit will detect.
Can we use this kit with cells that we already fixed?
It depends on the type of fixative used. The fixative we provide with the kit keeps the enzymes alive. If other fixative is used that kills the enzyme, then it can not be used with this kit.
What can be used as a positive control?
HeLa cells passage over 38 times can be used as a positive control.
Ater incubation, there seemed to have some crystals appearing in the wells. Why is that?
These crystals are salt crystals formed due to the solvent evaporation. Our recommendation is to keep the plate sealed when is left overnight. You may try to wash the staining with PBS, hopefully the crystal can get into PBS.
How many cells should be used in a 12-well plate?
2 – 5 Million cells dependent on the cell size
Customer wants to use rat and human SWAN cells with this assay. Laminin is necessary to adhere cells to TC plate. Would the laminin interfere with the assay?
No, laminin will not interfere with the assay. You are going to isolate the cells out of the plate for the staining. The cells would be washed with the PBS before staining. Therefore there would not be any traces of laminin left over.
I understand that this Senescence Detection Kit is designed for producing results that can be observed using a light microscope, but is it possible to read my results using a plate reader? If so, could you tell me what excitation and emissions wavelengths to use?Alternatively, could you provide some additional information about the type of dye used in this kit?
I am sorry but the colour development from this kit has to be analyzed using a microscope. It uses the X-gal staining system.
Can this kit be used with samples like bacteria, plants, drosophila, yeast etc?
We have optimized the kit with mammalian samples. However, theoretically these kits should work with samples from multiple species/sources. Since the optimal conditions depend on the sample type, the protocol has to be be adapted to fit the samples for efficient results. Please refer to this kit's citations to see what kind of samples have been used with this kit other than mammalian samples.
Can we use frozen samples with this assay?
Fresh samples are always preferred over frozen samples. However, frozen samples can also be used, provided, they were frozen right after isolation, were not freeze thawed multiple time (for which we recommend aliquoting the samples before freezing) and have been frozen for relatively short periods.
Can we use a different wavelength than recommended for the final analysis?
It is always recommended to use the exact recommended wavelength for the most efficient results. However, most plate readers have flexibility in their band width of detection in increments of +/- 10 nm. Depending on this flexibility range, you can deviate from the recommended wavelengths within limits.
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact your regional BioVision distributor.
What is the shelf life of this kit?
This kit is good for 12 months from the date of shipment in the unopened form when stored at the appropriate temperature and appropriate conditions. After opening and reconstitution, some of the components in this kit are good for 2 months at -20°C. Please refer to the datasheet for storage information and shelf life of each of the components.
Why are my standard curve values lower than those shown on the datasheet?
There are multiple factors which influence the signals like the incubation times, room temperature, handling etc. In general, to increase the value of the standards, you can increase the incubation time. As long as the standard curve is linear, it should be fine to use, since all of your samples will also be measured under the same conditions on this curve.
How do I normalize my samples against protein concentration
You can use a protein quantitation assay on the supernatants you get from cell/tissue lysates or with any other liquid sample in the assay buffer.
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Should I make a standard curve for every expt I do, or is one curve/kit enough?
Yes, I would strongly recommend you to do the standards every time you do the expt. There is always a chance that something was done differently that day and we do not want any conditions to differ between standards and samples.
Are the pH conditions used for this assay acidic in nature?
Yes, the pH condition used for this assay is acidic in nature.
Ok Hee Jeon, Senescence cell–associated extracellular vesicles serve as osteoarthritis disease and therapeutic markers. JCI Insight, April 2019; 30944259.
Su-Jeong Kim, GRSF1 is an age-related regulator of senescence. Sci Rep, April 2019;  30944385.
Fan Zhang, MYC and RAS are unable to cooperate in overcoming cellular senescence and apoptosis in normal human fibroblasts. Cell Cycle, Dec 2018;  30526305.
Cristina Guarducci, Cyclin E1 and Rb modulation as common events at time of resistance to palbociclib in hormone receptor-positive breast cancer. NPJ Breast Cancer., Nov 2018; 30511015.
Yi, Zhang et al. (2017) Effects of Nonsteroidal Anti-inflammatory Drugs on the Self-renewal Capacity of Blast Progenitors in Hematological Malignancies, J Environ Pathol Toxicol Oncol. 2017;36(1):15-27.
For more citations of this product click here