Home » ASK1 » ASK1 Proteins » Recombinant Human Active ASK1 (aa 649-946) Protein, CF
Novus Biologicals products are now on bio-techne.com

Recombinant Human Active ASK1 (aa 649-946) Protein, CF

 (1 Publication)
 See All Related
Submit a Review

Images

 
(
Enlarge)
The approximate molecular weight is 60 kDa and the average purity is 90%.
The approximate molecular weight is 60 kDa and the average purity is 90%.

Product Details

Summary
Reactivity HuSpecies Glossary
Applications Bioactivity
Format
Carrier-Free

Order Details

Novus Biologicals is part of Bio-Techne

Shop this product on bio-techne.com

Novus Biologicals, part of Bio-Techne, offers 100% Guarantee

Recombinant Human Active ASK1 (aa 649-946) Protein, CF Summary

Details of Functionality
The specific activity of ASK1 is typically 82-112 nmol/min/mg using a myelin basic protein (MBP) substrate.
Source
Spodoptera frugiperda, Sf 9 (baculovirus)-derived human ASK1 protein
aa 649-946
Accession #
N-terminal Sequence
Using an N-terminal GST tag
Protein/Peptide Type
Recombinant Proteins
Gene
MAP3K5
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.

Applications/Dilutions

Dilutions
  • Bioactivity
SDS-PAGE
60 kDa
Publications
Read Publication using
3575-KS in the following applications:

Packaging, Storage & Formulations

Storage
This product is stable at ≤ ‑70 °C for up to 1 year from the date of receipt. For optimal storage, aliquot into smaller quantities after centrifugation and store at recommended temperature. Avoid repeated freeze-thaw cycles.
Buffer
Supplied in 50 mM Tris HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, 10 mM glutathione, 0.1 mM EDTA, 0.1 mM PMSF, 25% glycerol.
Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane.
Assay Procedure
  • Active Kinase - Active ASK1 (0.1 μg/μL) diluted with Kinase Dilution Buffer and assayed as outlined in Figure 2. Note: These are suggested working dilutions. Optimal dilutions should be determined by each laboratory for each application.
  • Kinase Assay Buffer I, pH 7.2 - 25 mM MOPS, 12.5 mM beta -glycerolphosphate, 25 mM MgCl2, 5 mM EGTA, 2 mM EDTA. Add 0.25 mM DTT to the Kinase Assay Buffer prior to use.
  • Kinase Dilution Buffer, pH 7.2 - Kinase Assay Buffer I diluted at a 1:4 ratio (5-fold dilution) with 50 ng/μL BSA solution.
  • 10 mM ATP Stock Solution - Prepare the ATP Stock Solution by dissolving 55 mg of ATP in 10 mL of Kinase Assay Buffer I.
  • [33P]-ATP Assay Cocktail - Prepare 250 μM [33P]-ATP Assay Cocktail in a designated radioactive work area by combining 150 μL of 10 mM ATP Stock Solution, 100 μL of [33P]-ATP (1 mCi/100 μL), and 5.75 mL of Kinase Assay Buffer I.
  • Substrate - Myelin basic protein (MBP) diluted in distilled or deionized water to a final concentration of 1 mg/mL.
  1. Thaw the [33P]-ATP Assay Cocktail in a shielded container in a designated radioactive work area.
  2. Thaw the Active ASK1, Kinase Assay Buffer I, Substrate, and Kinase Dilution Buffer on ice.
  3. In a pre-cooled microfuge tube, add the following reaction components bringing the initial reaction volume up to 20 μL.
    a. Diluted Active ASK1: 10 μL
    b. Substrate (1 mg/mL Stock Solution): 5 μL
    c. Distilled water (2-8 °C): 5 μL
  4. Set up the blank control as outlined in step 3, excluding the addition of the substrate. Replace the substrate with an equal volume of distilled or deionized water.
  5. Initiate the reaction by the addition of 5 μL [33P]-ATP Assay Cocktail, bringing the final volume up to 25 μL. Incubate the mixture in a water bath at 30 °C for 15 minutes.
  6. After the 15 minute incubation period, terminate the reaction by spotting 20 μL of the reaction mixture onto individual pre-cut strips of phosphocellulose P81 paper.
  7. Air dry the pre-cut P81 strip and sequentially wash in a 1% phosphoric acid solution (dilute 10 mL of phosphoric acid and make a 1 liter solution with distilled or deionized water) with constant gentle stirring. It is recommended that the strips be washed a total of three times for approximately 10 minutes each.
  8. Count the radioactivity on the P81 paper in the presence of scintillation fluid in a scintillation counter.
  9. Determine the corrected cpm by removing the blank control value (see step 4) for each sample and calculate the kinase specific activity as outlined below:


    Calculation of [33P]-ATP Specific Activity (SA) (cpm/pmol)
    Specific Activity (SA) = cpm for 5 μL [33P]-ATP/pmole of ATP (in 5 μL of a 250 μM ATP stock solution; i.e. 1250 pmol)

    Calculation of Kinase Specific Activity (SA) (pmol/minutes/μg or nmol/minutes/mg)
    Corrected cpm from reaction / [(SA of 33P-ATP in cpm/pmol) x (Reaction time in minutes) x (Enzyme amount in μg or mg)] x [(Reaction volume) / (Spot Volume)]

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Recombinant Human Active ASK1 (aa 649-946) Protein, CF

  • Apoptosis signal-regulating kinase 1
  • ASK1
  • ASK-1
  • ASK1MEKK 5
  • EC 2.7.11
  • MAP/ERK kinase kinase 5
  • MAP3K5
  • MAPK/ERK kinase kinase 5
  • MAPKKK5EC 2.7.11.25
  • MEK kinase 5
  • MEKK5
  • MEKK5apoptosis signal regulating kinase 1
  • mitogen-activated protein kinase kinase kinase 5

Background

ASK1, also known as MEKK5 or MAPKKK5, activates MKK3, MKK4 (SEK1), and MKK6. Over-expression of ASK1 induces apoptotic cell death and is activated in cells treated with tumor necrosis factor alpha (TNF ) (1). ASK1 interacts with members of the TRAF family and is activated by TRAF2 in the TNF-signaling pathway. After activation by TRAF2, ASK1 activates MKK4, which in turn activates JNK. Thus, ASK1 is a mediator of TRAF2-induced JNK activation (2).
  1. Ichijo, H. et al. (1997) Science 275:90.
  2. Nishitoh, H. et al. (1998) Mol. Cell 2:389.

Customers Who Viewed This Item Also Viewed...

JNK1 Antibody
(228601) [Unconjugate
MAB17761
Western Blot JNK1 Antibody (228601) [Unconjugated]
Western Blot JNK1 Antibody (228601) [Unconjugated]
Immunocytochemistry JNK1 Antibody (228601) [Unconjugated]
(1 Review)
(3 Publications)
Species: Hu, Mu, Rt
Applications: ICC, WB
TNF-alpha
[Unconjugated]
210-TA
SDS-PAGE TNF-alpha [Unconjugated]
Bioactivity TNF-alpha [Unconjugated]
SEC-MALS TNF-alpha [Unconjugated]
(3 Reviews)
(821 Publications)
Species: Hu
Applications: BA
ERK2 Antibody
[Unconjugated]
AF1230
Western Blot ERK2 Antibody [Unconjugated]
Immunohistochemistry ERK2 Antibody [Unconjugated]
Knockout Validated ERK2 Antibody [Unconjugated]
(3 Publications)
Species: Hu, Mu, Rt
Applications: IHC, KO, WB
p38 alpha Antibody
[Unconjugated]
AF8691
Simple Western p38 alpha Antibody [Unconjugated]
Western Blot p38 alpha Antibody [Unconjugated]
Immunohistochemistry p38 alpha Antibody [Unconjugated]
(1 Review)
(11 Publications)
Species: Hu, Mu, Rt
Applications: IHC, KO, Simple Western, WB
Thioredoxin-1
Antibody - BSA Free
NBP2-52983
Flow Cytometry Thioredoxin-1 Antibody - BSA Free
Immunocytochemistry/ Immunofluorescence Thioredoxin-1 Antibody - BSA Free
Immunocytochemistry/ Immunofluorescence Thioredoxin-1 Antibody - BSA Free
(1 Publication)
Species: Hu, Mu
Applications: Flow, ICC/IF, IHC, IHC-P, WB
POLDIP2 Antibody
NBP1-68874
Western Blot POLDIP2 Antibody
Western Blot POLDIP2 Antibody
(1 Publication)
Species: Hu, Mu, Rt
Applications: PEP-ELISA, WB
Crk Antibody
(3G11E8) - BSA Free
NBP2-37568
ELISA Crk Antibody (3G11E8) - BSA Free
Western Blot Crk Antibody (3G11E8) - BSA Free
Western Blot Crk Antibody (3G11E8) - BSA Free
Species: Hu
Applications: ELISA, Flow, ICC/IF, IHC, IHC-P, WB
GRAP2 Antibody
(OTI1G2)
NBP2-00764
Flow Cytometry GRAP2 Antibody (OTI1G2)
Flow Cytometry GRAP2 Antibody (OTI1G2)
Western Blot GRAP2 Antibody (OTI1G2)
Species: Hu, Mu, Rt
Applications: Flow, IHC, IHC-P, WB
AHSA1 Antibody
(1A2-A8)
H00010598-M01
Western Blot AHSA1 Antibody (1A2-A8)
Sandwich ELISA AHSA1 Antibody (1A2-A8)
(6 Publications)
Species: Hu, Mu, Rt
Applications: ELISA, ICC/IF, S-ELISA, WB
AIMP2 Antibody
NBP1-81575
Immunohistochemistry-Paraffin AIMP2 Antibody
Immunohistochemistry-Paraffin AIMP2 Antibody
Immunohistochemistry-Paraffin AIMP2 Antibody
Species: Hu, Mu, Rt
Applications: ICC/IF, IHC, IHC-P, WB
c-jun [p Ser63]
Antibody (1018Y) [Unc ...
MAB8930
Western Blot c-jun [p Ser63] Antibody (1018Y) [Unconjugated]
Immunocytochemistry c-jun [p Ser63] Antibody (1018Y) [Unconjugated]
Simple Western c-jun [p Ser63] Antibody (1018Y) [Unconjugated]
(1 Review)
(3 Publications)
Species: Hu
Applications: ICC, Simple Western, WB
MKK4/MEK4 Antibody
(OTI8A8)
NBP1-47839
Flow Cytometry MKK4/MEK4 Antibody (OTI8A8)
Immunocytochemistry/ Immunofluorescence MKK4/MEK4 Antibody (OTI8A8)
Immunocytochemistry/ Immunofluorescence MKK4/MEK4 Antibody (OTI8A8)
(1 Review)
Species: Ca, Hu, Mu, Rt
Applications: Flow, ICC/IF, IHC, IHC-P, IP, WB
SGSM3 Antibody
NBP2-13304
Immunohistochemistry-Paraffin SGSM3 Antibody
Immunocytochemistry/ Immunofluorescence SGSM3 Antibody
Western Blot SGSM3 Antibody
Species: Hu
Applications: ICC/IF, IHC, IHC-P, WB
TRAF-2 Antibody
NB100-56173
Immunocytochemistry/ Immunofluorescence TRAF-2 Antibody
Immunohistochemistry TRAF-2 Antibody
Immunohistochemistry-Paraffin TRAF-2 Antibody
(4 Publications)
Species: Hu
Applications: Flow, ICC/IF, IHC, IHC-P, IP, WB
MKK3/MEK3 Antibody
(2E12D11) - BSA Free
NBP2-52508
ELISA MKK3/MEK3 Antibody (2E12D11) - BSA Free
Flow Cytometry MKK3/MEK3 Antibody (2E12D11) - BSA Free
Immunofluorescence MKK3/MEK3 Antibody (2E12D11) - BSA Free
Species: Hu
Applications: CyTOF-ready, ELISA, Flow, ICC, ICC/IF, IHC, IHC-P, WB
VAC14 Antibody
H00055697-B01P
Immunocytochemistry/ Immunofluorescence VAC14 Antibody
Western Blot VAC14 Antibody
Western Blot VAC14 Antibody
Species: Hu
Applications: ICC/IF, WB
Caspase-3 Antibody
[Unconjugated] - Acti ...
AF835
Immunocytochemistry Caspase-3 Antibody [Unconjugated] - Active
Immunohistochemistry Caspase-3 Antibody [Unconjugated] - Active
Immunocytochemistry Caspase-3 Antibody [Unconjugated] - Active
(7 Reviews)
(389 Publications)
Species: Hu, Mu
Applications: ICC, IHC
AKT [p Ser473]
Antibody [Unconjugate ...
AF887
Western Blot AKT [p Ser473] Antibody [Unconjugated] - Pan Specific
Simple Western AKT [p Ser473] Antibody [Unconjugated] - Pan Specific
Intracellular Staining by Flow Cytometry AKT [p Ser473] Antibody [Unconjugated] - Pan Specific
(50 Publications)
Species: Hu, Mu, Rt
Applications: CyTOF-ready, IHC, ICFlow, Simple Western, WB

Publications for ASK1 (3575-KS)(1)

We have publications tested in 1 confirmed species: Human.

We have publications tested in 1 application: Bioassay.


Filter By Application
Bioassay
(1)
All Applications
Filter By Species
Human
(1)
All Species

Reviews for ASK1 (3575-KS) (0)

There are no reviews for ASK1 (3575-KS). By submitting a review you will receive an Amazon e-Gift Card or Novus Product Discount.
  • Review with no image -- $10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen
  • Review with an image -- $25/€18/£15/$25 CAD/¥150 Yuan/¥2500 Yen

FAQs for ASK1 (3575-KS) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Additional ASK1 Products

Blogs on ASK1.

Check out the latest blog posts on ASK1.
Liver ASK1 activates autophagy to protect against hepatic fat accumulation, non-alcoholic steatohepatitis and fibrosis
By Jamshed Arslan, Pharm. D., PhD. The most common chronic liver disorder worldwide is non-alcoholic fatty liver disease (NAFLD). This obesity-linked disorder can manifest as hepatic fat accumulation (steatosis) wit...  Read full blog post.
IRE1 alpha dependent apoptotic-signaling pathway
Despite in depth characterization of the role of IRE1 alpha (inositol-requiring enzyme 1 alpha) in activating the unfolded protein response (UPR) in the ER - little is known about the molecular mechanisms by which this ER protein has shown to reg...  Read full blog post.
Read our latest blog and use the new citation tool on bio-techne.com

Customers Who Bought This Also Bought

Caspase‑3 was detected in immersion fixed anti-FAS treated Jurkat human acute T cell leukemia cell line using 0.3 µg/mL Human/Mouse Active Caspase‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF835) for 3 hours at room temperature. Cells were stained (red) and counterstained (green). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Caspase-3 Antibody [Unconjugated] - Active
AF835
1 µg/lane of Recombinant Human TNF-alpha was resolved by SDS-PAGE with silver staining, under reducing (R) conditions, showing a band at 17 kDa.
TNF-alpha [Unconjugated]
210-TA-005

Contact Information

Product PDFs

  PDF SDS
  PDF Datasheet
  Request CofA

Calculators

Concentration Calculator

The concentration calculator allows you to quickly calculate the volume, mass or concentration of your vial. Simply enter your mass, volume, or concentration values for your reagent and the calculator will determine the rest.

=
÷

Review this Product

Be the first to review our Recombinant Human Active ASK1 (aa 649-946) Protein, CF and receive a gift card or discount.


Bioinformatics

Gene Symbol MAP3K5
Uniprot