products | cell biology and antibodies | Apoptosis, Signal Transduction, Cytotoxicity and Cell Proliferation

Cell Biology and Antibodies

Apoptosis, Signal Transduction, Cytotoxicity and Cell Proliferation

IL-18, Human and Mouse - Recombinant Cytokine, Antibodies and ELISA Kits

Introduction:

Interleukin 18 (IL-18) is an 18 kDa cytokine which is identified as a co-stimulatory factor for production of interferon-gamma in response to toxic shock. These new and unique IL-18 products are used for:

• determination of IL-18 concentration (ELISA)
  
• detection and neutralization of IL-18 (antibodies)
  
• analysis of IL-18 (recombinant protein)
  

Also available: Various human cytokine ELISA kits and monoclonal antibodies against IL-1, IL-2, IL-3R, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12p40 and IL-13

Anti-Fas Antibodies, Human and Mouse for Induction of Apoptosis

We offer different monoclonal anti-Fas antibodies including two which can directly induce apoptosis of cells. The corresponding cell surface antigen "Fas" belongs to the tumor necrosis factor receptor (TNFR) super family. For Western blots, immunohistochemistry and flow cytometry.

Also available: sFas (S) ELISA Kit and sFas Ligand ELISA Kit

Caspase Protease Assay Kits

Rapid detection of early stages of apoptosis

Product Description:

The activation of ICE-family proteases (caspases) initiates apoptosis in mammalian cells by degrading critical cellular proteins associated with cell structure, DNA repair and transcriptional regulation. The Caspase Protease Assay Kits provide a simple and convenient assay for the activity of several types of caspases, based on their recognition of specific amino acid sequences. Comparison of the readings from an apoptotic sample with an uninduced control allows determination of the fold-increase in caspase activity. Unlike comparable assays that require purified protein, the Caspase Protease Assay Kits can be used with crude cell lysates, offering considerable time savings. Two types of kits are available: fluorescent and colorimetric.

Caspase-1, Caspase-2, Caspase-3, Caspase-5, Caspase-6, Caspase-8, Caspase-9 and Caspase-10 Colorimetric and Fluorometric Protease Assay Kits are available.

EnzChek^® Caspase-3 Assay Kit

Sensitive, fluorescence-based apoptosis detection

Product Description:

Caspase-3 is the key effector in the apoptosis pathway, amplifying the signal from initiator caspases (such as caspase-8) and signifying full commitment to cellular diassembly. In addition to cleaving other caspases in the enzyme cascade, caspase-3 has been shown to cleave poly(ADP-ribose) polymerase (PARP), DNA-dependent protein kinase, protein kinase Cd and actin. We offer a selection of fluorogenic substrates containing the caspase-3 recognition site Asp-Glu-Val-Asp (DEVD) which are the main components in our simple-to-use Caspase-3 Assay Kits.

Annexin V Apoptosis Detection Kits

Early detection of apoptosis in living cells

Product Description:

During the early stages of apoptosis, cell membranes lose their phospholipid symmetry and expose phosphatidylserine (PS) at the cell surface. Annexin V, a calcium-dependent phospholipid-binding protein, has a high affinity for PS. The Apoptosis Detection Kits use Annexin V conjugated to various markers or chromophores for convenient detection of apoptotic cells. They can detect apoptosis much earlier than conventional assays which are based on detecting morphological changes that occur later than PS translocation. Since no fixation procedures are required, these kits can be used to study apoptosis directly in living cells.

PS translocation also occurs in cell necrosis; however, in this case the cell membrane loses its integrity. Thus, necrotic cells can be stained with propidium iodide as well as Annexin V conjugate while apoptotic cells will be stained only with Annexin V conjugate. The kits contain the appropriate Annexin V conjugate, binding buffer, and propidium iodide and can be used for detecting apoptosis by e.g. fluorescence microscopy or flow cytometry.

Available Kits from AG Scientific: Annexin V-Biotin, Annexin V-APC and Annexin V-FITC Apoptosis Detection Kits.

MEBSTAIN Apoptosis Kit Direct

In situ detection of apoptosis by the TUNEL method

Product Description:

The MEBSTAIN Apoptosis Kit Direct provides a sensitive and rapid method for detecting apoptosis. The kit is suitable for use in histochemistry, cytospin preparations, and flow cytometry. The protocol is based on the TdT-mediated dUTP nick end-labeling (TUNEL) method which labels the 3´-OH ends of DNA fragments generated by apoptosis. The MEBSTAIN Apoptosis Kit Direct uses fluorescein-dUTP as the label, allowing sensitive detection of DNA fragmentation by fluorescence microscopy. This direct detection method eliminates the need for a secondary detection step with antibody conjugates which is required with conventional biotin-based systems.

Also available: APO-BrdU™ TUNEL Assay Kit which uses BrdUTP 5-bromo-2'-deoxyuridine 5'-triphosphate) to label the DNA strand breaks and Alexa Fluor^® 488 dye-labeled anti-BrdU antibodies for the final detection of the BrdU incorporated into the DNA.

MitoCapture™ Apoptosis Detection Kit

Detects changes in the mitochondrial membrane potential

Product Description:

Disruption of the mitochondrial membrane potential is one of the earliest intracellular events that occur following induction of apoptosis. The MitoCapture™ Apoptosis Detection Kit provides a fluorescent-based method for distinguishing between healthy and apoptotic cells by detecting the changes in the mitochondrial membrane potential.

Using our MitoTracker^® Red CMXRos is an additional, very quick and reliable method for detecting apoptosis-induced mitochondrial membrane depolarization by flow cytometry. This probe accumulates readily in active mitochondria but only to a lesser degree in mitochondria with depolarized membranes. The relative brightness of the mitochondrial staining therefore provides a way to distinguish healthy from apoptotic cells.

ApopLadder Ex™

Apoptotic DNA Fragment Extraction Kit

Features:

• ready-to-use: no reagent preparation necessary
• sensitive detection of apoptosis DNA fragments
• high specificity: selective extraction of fragmented DNA with less extraction of intact chromatin DNA
• rapid: results after 2.5 hours
• quantitative: quantification of DNA fragments can be done by fluorescent staining
• safe: no phenol and chloroform required
  
  

Product Description:

Apoptosis is associated with the fragmentation of chromosomal DNA into approximately 180 bp nucleosome units. ApopLadder Ex™ is designed to selectively extract fragmented DNA from apoptotic cells. The kit enables the extraction of even small DNA fragments by avoiding the extraction of intact chromatin. This results in a highly sensitive detection and quantitation of fragmented DNA which can be visualized as a DNA ladder by electrophoresis and can be quantified by fluorescent staining.
The kit contains a lysis buffer with a special detergent, enzyme, DNA precipitant and 6x loading buffer. The following procedures can be performed in approximately 2.5 hours:

1. cell lysis
2. centrifugation
3. enzymatic degradation of proteins and RNA
4. precipitation of DNA fragments
   

Also available:

• TUNEL assay to detect apoptotic cells using ChromaTide™ BODIPY^® FL-14-dUTP. This direct TUNEL assay has been reported to be significantly more sensitive than the indirect TUNEL method using biotin or digoxygenin. Furthermore, the narrow fluorescence emission of the BODIPY^® FL dye (absorption and emission peaks at 505 nm and 515 nm, respectively) makes it compatible with multiparameter labeling.
  
  
• Quick Cell Proliferation Assay Kit: Requires less than 90 minutes to prepare DNA in a single tube without the need of extraction or using columns.
  
  
• ApoPrimer Set (Bcl-2 family) Detects mRNA of 7 typical members of the human Bcl-2 family and beta-actin mRNA by RT-PCR.
  

LIVE/DEAD^® Viability/Cytotoxicity Kit

A fast and reliable viability test for eukaryotic cells

Features:

• sensitive and accurate indicator of cytotoxic events - suitable for HTS of cytotoxic agents
• more reliable than trypan blue
• easy and quick analysis with all fluorescence detection systems
• faster, cheaper and safer than ⁵¹Cr-release assays
  
  

Product Description:

The fluorescent probes included in this kit, calcein AM and ethidium homodimer-1, provide a precise, general-purpose assay of cytotoxic events: live cells cleave membrane-permeant AM to yield cytoplasmic green fluorescence; membrane-impermeant ethidium homodimer-1 labels nucleic acids of membrane-compromized cells with red fluorescence. The ease, reliability and low cost of this kit makes it economical for high-throughput screening of cytotoxic agents.

Also available: LIVE/DEAD^® Cell-Mediated Cytotoxicity Kit, LIVE/DEAD^® Reduced Biohazard Viability/Cytotoxicity Kit, LIVE/DEAD^® Sperm Viability Kit, LIVE/DEAD^® Yeast Viability Kit and the LIVE/DEAD BacLight Bacterial Viability Kit.

LDH Cytotoxicity Detection Kit

Cell death measurement and analysis of cytotoxic compounds

Product Description:

Many widely-used methods for measuring cell viability are based on the uptake or exclusion of dyes, such as trypan blue or eosin Y. These methods cannot process large numbers of samples and are not quantitative. Other methods are based on the measurement of released radioactive isotopes or fluorescent dyes from prelabeled target cells. However, these cells must be prelabeled and can spontaneously release the labels.
Instead, the LDH Cytotoxicity Detection Kit is based on the measurement of lactate dehydrogenase (LDH) released from damaged cells. As it is released into the cell culture supernatant, LDH participates in a coupled reaction converting a yellow tetrazolium salt to red formazan. The amount of enzyme activity correlates to the number of damaged cells. The entire procedure, including cell growth and color measurement, may be performed in a single multiwell plate.

For details click here.

CyQUANT^® Cell Proliferation Assay Kit

Assessing cell proliferation accurately and fast

Product Description:

This kit provides a fluorescence-based method that is sensitive enough to replace tedious and costly ³H-thymidine incorporation assays. Although it does not directly measure DNA synthesis, the assay does measure the result of DNA synthesis — the fluorescence intensity of the CyQUANT^® GR dye increases in proportion to the quantity of nucleic acid present (i.e. the number of cells). In a fluorescence microplate reader, this assay has a linear range from 50 to 50,000 cells using a single dye concentration, making it easily adaptable for automated high-throughput experiments. For added convenience, samples may be frozen indefinitely before analysis. This assay is particularly useful for screening various compounds that affect cell proliferation, such as tumor suppressors and growth factors.

ABSOLUTE-S™ SBIP Cell Proliferation Assay Kit

Improved assay based on the TUNEL method

Product Description:

Many conventional BrdU-based protocols for assaying cell proliferation require DNA denaturation to expose the incorporated BrdU. However, such harsh treatment often makes it difficult to perform additional staining, because antigens and other cellular structures are not well preserved. In contrast, the ABSOLUTE-S™ SBIP Cell Proliferation Assay Kit, offered in conjunction with Phoenix Flow Systems, Inc., uses the SBIP (Strand Breaks Induced by Photolysis) methodology which better preserves cellular features and antigen sites. In the ABSOLUTE-S™ assay, cells are first incubated in the presence of BrdU which is incorporated into cellular DNA during replication. BrdU incorporation results in sensitization to photolysis; UV light is used to induce breaks at these sites. Additional BrdU is added at the break sites using TUNEL. Finally, BrdU is detected using an Alexa Fluor^® 488 dye-labeled anti-BrdU antibody (included). This kit is compatible with flow cytometry and fluorescence microscopy and contains fixed control cells to assist in the interpretation of the results.

Universal Tyrosine Kinase and Tyrosine Phosphatase Assay Kits

Measure PTK and PTP activity fast and specifically

Features:

• safety: no radioactive isotopes are required
• accuracy: low background and no cross reactions with Protein Ser/Thr Kinase or Protein Ser/Thr Phosphatase
• sensitivity is comparable to radioactive tests
• ease of use: preparation of solutions is very easy, kinase or tyrosine phosphatase standards are included in the kits
• flexibility: synthesized peptides are designed to cover a wide spectrum for various PTKs and PTPs; combining a specific antibody enables to specifically measure PTK or PTP activity
  
  

Product Description:

The Universal Tyrosine Kinase Assay Kit enables a quick and specific measurement of Protein Tyrosine Kinase activity in a wide range with non-RI chemicals. It is useful for analysis of PTK activity regulation by using the recombinant PTK and for the in vitro screening of PTK inhibitors.

Protein Tyrosine Phosphatase (PTP) mediating protein dephosphorylation keeps the balance of those protein activities that are involved in cell proliferation and differentiation. Like PTK, PTP is classified into two groups: 1) membrane receptor PTPs and 2) PTPs, located in the cytoplasm. As PTP acts on a wide range of substrate, it is also classified by its substrates into several groups. The Universal Tyrosine Phosphatase Assay Kit enables a fast and specific in vitro measurement of PTP activity using a non-RI method. As this kit utilizes the mono-peptide substrate having a broad PTP specificity, it is useful to measure the activity of various PTPs. It can also be used for screening of selective PTP inhibitors, not only for screening of PTP itself.

Premix WST-1 Cell Proliferation Assay

Measurement of cell proliferation or cell viability

Features:

• safety: no radioactive isotopes and volatile organic solvents are required
• accuracy: absorbance strongly correlates to the number of viable cells
• sensitivity: more sensitive than using MTT
• ease of use and and short reaction time
• flexibility: microtiter plates can be read and returned several times to the incubator for further color development
  
  

Product Description:

For quantifying cell proliferation or cell viability, a new non-radioactive method has recently been developed which measures the number of viable cells via colorimetric detection of cleaved tetrazolium salts that are added into the medium. Washing and collection of cells are not required, and all procedures, from small-scale culture to data analysis with an ELISA reader, can be carried out in the same microtiter plate.
The PreMix WST-1 is based on the cleavage of tetrazolium salts (WST-1) to soluble formazan by a mitochondrial dehydrogenase -the succinate-tetrazolium reductase - which exists in the mitochondrial respiratory chain and is active only in viable cells. Thus, total activity of this enzyme in a sample rises with the increase of viable cells and the quantity of produced formazan dye which can be measured using an ELISA reader is directly proportional to the number of metabolically active cells in the medium.

For details click here.