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Cell biology assays

Signal transduction dominant negative vectors

Investigate the activity and interactions of key signal transduction pathways with our dominant-negative vector sets. Each vector sets includes a vector that expresses a strong wild-type protein and one that provides a dominant negative, nonfunctional version—allowing you to either activate or "knock down" the pathway. To set up a complete assay system, you can co-express these vectors with an appropriate cis-acting reporter vector to measure the activation of a signal transduction pathway and quantify differences in activation.

p53

The p53 Dominant-Negative Vector Set lets you study p53 and related signaling pathways. These vectors constitutively express high levels of wild-type p53 or its dominant-negative mutant. To determine the effects of p53 expression on a particular cell line, simply transfect with the vector containing the gene for wild-type p53, and examine morphology or look for changes in the expression of other pathway proteins.

IkappaBalpha (IKBA)

The IkappaBalpha Dominant-Negative Vector Set provides a convenient way to examine NFkappaB regulation by manipulating its inhibitor, IkappaBalpha, which normally keeps NFkappaB inactive and sequestered in the cytosol. This vector set gives you the ability to compare the effects of overexpressed IkappaBalpha (phosphorylated specifically by your agent) to those of IkappaBalphaM (unable to be phosphorylated under any conditions). By combining these vectors with cis-acting NFkappaB vectors, you have a complete assay system to study and measure activation of the NFkappaB pathway.

CREB

The CREB Dominant-Negative Vector Set includes a constitutively active mutant and two different dominant-negative mutants which allow you to block CREB signaling by separate mechanisms in order to determine which genes are activated by CREB (cAMP response element-binding protein)—a transcription factor that acts downstream in the protein kinase A (PKA) signaling pathway.

Ras & Raf

The Ras Dominant-Negative Vector Set lets you study Ras, a GTP-binding protein that relays signals affecting many cellular processes, including human tumor development (Malumbres and Pellicer, 1998). We also offer a vector set for studying Raf, the most highly characterized downstream effector of Ras. You can combine the Ras or Raf vector sets with our pathway profiling systems to easily monitor cross-talk between different signaling pathways.

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Cat. #	Product	Size	Price
631927	Kinase Expression Vector Set	Each	USD $645.00
This Kinase Expression Vector Set contains three plasmids that constitutively express one of the following kinases: MEK1 (human), MEKK1 (mouse), and PKA (mouse). These plasmids can be used in conjunction with our In Vivo Kinase Assay Kits or for other expression studies involving these kinases. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription.**
631926	Raf Dominant-Negative Vector Set	Each	USD $645.00
Set of three vectors for studying Raf or MAPK signaling pathways: the pCMV-Raf-1 Vector expresses the wild-type Raf protein, the pCMV-RafCAAX expresses a constitutively active form of the Raf protein, and the pCMV-RafS621A Vector expresses a mutated form of Ras with a Ser-to-Ala mutation at residue 621. These vectors can be used to study Raf-related signal transduction pathways in mammalian cells. Each gene is expressed from the constitutive CMV promoter. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription. Back Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set (Cat. # 631923).** HEK 293 cells were transiently transfected with the indicated plasmids. After 16 hr, fresh medium was added to the cells along with TNF to induce the NFκB pathway. SEAP activity was assayed 6 hr later to determine NFκB activity. Back 631926: Raf Dominant-Negative Vector Set
631925	CREB Dominant-Negative Vector Set	Each	USD $645.00
Set of three vectors for studying CREB-related signaling pathways. The pCMV-CREB Vector expresses the wild-type CREB protein. pCMVCREB133 Vector encodes a mutated form of CREB, blocking cAMP-induced transcription and preventing CREB phosphorylation. pCMVKCREB also encodes a mutated form of CREB, blocking cAMP-induced transcription. Each gene is expressed from the constitutive CMV promoter. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription. Back Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set (Cat. # 631923).** HEK 293 cells were transiently transfected with the indicated plasmids. After 16 hr, fresh medium was added to the cells along with TNF to induce the NFκB pathway. SEAP activity was assayed 6 hr later to determine NFκB activity. Back 631925: CREB Dominant-Negative Vector Set
631924	Ras Dominant-Negative Vector Set	Each	USD $645.00
Set of three vectors for studying Ras or MAPK signaling pathways. The pCMV-Ras Vector expresses the wild-type Ha-ras protein. The pCMV-RasN17 Vector contains a mutated form of Ras with a serine-to-asparagine mutation at residue 17. To constitutively express Ras, pCMV-RasV12 contains a mutated form of Ras with a glycine-to-valine mutation at residue 12. These vectors can be used to study Ras-related signal transduction pathways in mammalian cells. Each gene is expressed from the constitutive CMV promoter. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription. Back Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set (Cat. # 631923).** HEK 293 cells were transiently transfected with the indicated plasmids. After 16 hr, fresh medium was added to the cells along with TNF to induce the NFκB pathway. SEAP activity was assayed 6 hr later to determine NFκB activity. Back 631924: Ras Dominant-Negative Vector Set
631923	IkappaBalpha Dominant-Negative Vector Set	Each	USD $645.00
Set of two vectors for studying NFκB and related signaling pathways. The pCMV-IκBα Vector expresses the wild-type IκBα protein. The pCMV-IκBαM Vector contains a mutated form of IκBα with a serine-to-alanine mutation at residues 32 and 36. This mutated form does not undergo signal-induced phosphorylation, and thus, remains bound to NFκB. Both genes are expressed from the constitutive CMV promoter. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription. Back Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set (Cat. # 631923).** HEK 293 cells were transiently transfected with the indicated plasmids. After 16 hr, fresh medium was added to the cells along with TNF to induce the NFκB pathway. SEAP activity was assayed 6 hr later to determine NFκB activity. Back 631923: IkappaBalpha Dominant-Negative Vector Set
631922	p53 Dominant-Negative Vector Set	Each	USD $645.00
Set of two vectors for studying p53 and related signaling pathways. pCMV-p53 expresses the wild-type p53 tumor suppressor protein. pCMV-p53mt135 contains the gene encoding a dominant-negative mutation of p53 that blocks normal p53 activity. Both genes are expressed from the constitutive CMV promoter. Use these vectors in mammalian systems for studying p53 function in apoptosis, cell growth, and DNA damage repair. Notice to purchaser Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Documents Components Image Data Back Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription *Signal transduction kits are available for in vivo* studies of signal transduction—from signaling cascades to gene transcription. Back Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set (Cat. # 631923).** HEK 293 cells were transiently transfected with the indicated plasmids. After 16 hr, fresh medium was added to the cells along with TNF to induce the NFκB pathway. SEAP activity was assayed 6 hr later to determine NFκB activity. Back 631922: p53 Dominant-Negative Vector Set

Overview

Simplify the complexities of signal transduction in vivo
Dissect the network of signal transduction pathways
Screen for compounds that activate key pathways
Combine with cis reporter vectors to create a complete assay system

More Information

Applications

Measure and quantify signal transduction pathway activity
Activation and knockdown studies

References

Malumbres, M. & Pellicer, A. RAS pathways to cell cycle control and cell transformation. Front. Biosci. 3:d887–912 (1998).

Additional product information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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Signal transduction dominant negative vectors

p53

IkappaBalpha (IKBA)

CREB

Ras & Raf

Notice to purchaser

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Notice to purchaser

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set

631926: Raf Dominant-Negative Vector Set

Notice to purchaser

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set

631925: CREB Dominant-Negative Vector Set

Notice to purchaser

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set

631924: Ras Dominant-Negative Vector Set

Notice to purchaser

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set

631923: IkappaBalpha Dominant-Negative Vector Set

Notice to purchaser

Signal transduction kits are available for in vivo studies of signal transduction—from signaling cascades to gene transcription

Inhibition of NFκB activity using pCMV-IκBα and pCMVκIαBM from the IκB Dominant-Negative Vector Set

631922: p53 Dominant-Negative Vector Set

Overview

More Information

Applications

References

Additional product information