GeneTex
United States (US)

ASK1 (phospho Ser83) antibody

ASK1-phospho-Ser83-antibody_WB_GTX14380-1_18121409_561.jpg
ASK1-phospho-Ser83-antibody_WB_GTX14380-2_18121409_343.jpg
ASK1-phospho-Ser83-antibody_ELISA_GTX14380-1_18121409_617.jpg
ASK1-phospho-Ser83-antibody_IHC_GTX14380-1_18121409_797.jpg

Cat No. GTX14380

Host Rabbit
Clonality Polyclonal
Isotype IgG
Application WB, IHC-P, ELISA
Reactivity Human
APPLICATION

Application Note

For WB: Use at a dilution of 1:500-1:1000. Predicted molecular weight: 155 kDa. For IHC-P: Use at a dilution of 1:50-1:1000. Optimal dilutions/concentrations should be determined by the end user.

Calculated MW

155 kDa. ( Note )

Specificity/Sensitivity

ASK1 (phospho-Ser83) antibody detects endogenous levels of ASK1 only when phosphorylated at serine 83.
PROPERTIES

Form

Liquid

Buffer

Phosphate buffered saline, pH 7.3, 50% glycerol and 150mM NaCl containing 0.02% sodium azide

Storage

Store as concentrated solution. Centrifuge briefly prior to opening vial. For short-term storage (1-2 weeks), store at 4ºC. For long-term storage, aliquot and store at -20ºC or below. Avoid multiple freeze-thaw cycles.

Concentration

1 mg/ml (Please refer to the vial label for the specific concentration.)

Antigen Species

Human

Immunogen

The antiserum was produced against synthesized phosphopeptide derived from human ASK1 around the phosphorylation site of serine 83 (G-S-S^P-V-G).

Purification

Affinity purified with antigen
From polyclonal serum

Conjugation

Unconjugated

Note

For laboratory use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
TARGET

Synonyms

Mitogen-Activated Protein Kinase Kinase Kinase 5,Ask1,Mapkkk5,Mekk5,Map3K5

Cellular Localization

Cytoplasmic

Background

Mitogen-activated protein (MAP) kinase cascades are activated in response to various extracellular stimuli including cytokines, growth factors and environmental stresses. A novel MAP kinase kinase kinase (MAPKKK) was recently identified and designated ASK1 (for apoptosis signal-regulating kinase 1) and MAPKKK5. ASK1 activates two different subgroups of MAPKK, MKK4 and MKK6, which in turn activate c-Jun N-terminal kinase (JNK) and p38 MAP kinase, respectively. ASK1/MAPKKK5 is activated by TNFR and Fas through the interaction with members of the TRAF family and Fas-associated protein Daxx. Overexpression of ASK1 induces apoptotic cell death, and a catalytically inactive form of ASK1 inhibits TNF-a induced apoptosis. ASK1 is expressed in variety of human and mouse tissues.

Database

Research Area

DATA IMAGES
ASK1-phospho-Ser83-antibody_WB_GTX14380-1_18121409_561.jpg

GTX14380 WB Image

Western blot of anti-pS83 ASK1 antibodies (GTX14380) shows specificity for phosphorylated human ASK1. Anti-pS83 (aa 76-87) antibody was tested by western blot against Cos-7 cell lysates after transient transfection with 1) vector only, 2) recombinant HA-ASK1, and 3) recombinant human HA-ASK1 where S83 was substituted with an alanine residue. Cells were lysed 24 h post-transfection in 200 ?L of 1x SDS-sample buffer, heated at 96?C for 5', and vortexed for 30 sec. Samples (10 uL each) were separated on a 12% SDS-PAGE gel and transferred to PVDF (Millipore) followed by blocking for 45' using TTBS supplemented with 5% non-fat dry milk. All incubations were performed at room temperature. In panel a) all samples were incubated with anti-HA antibody. This blot demonstrates both recombinant transfections express rASK1. In panel b) all samples were incubated with anti-pS83 ASK1. Lane 2 shows strong specific staining of ASK1. Lane 3, where S83 was replaced with alanine, shows greatly diminished staining. In panel c) all samples were incubated with anti-pS83 ASK1 antibody as before except the antibody was pre-incubated with phospho peptide prior to membrane incubation. No staining is observed after phospho peptide blocking occurs.

ASK1-phospho-Ser83-antibody_WB_GTX14380-2_18121409_343.jpg

GTX14380 WB Image

Western blot analysis of extracts from K562 cells using ASK1 (phospho-Ser83) antibody (GTX14380)

ASK1-phospho-Ser83-antibody_ELISA_GTX14380-1_18121409_617.jpg

GTX14380 ELISA Image

ELISA results of purified polyclonal anti-pS83 ASK-1 (aa 76-87) antibody (GTX14380) tested against BSA conjugates of non-phospho and phospho forms of immunizing peptide. Each well was coated with 0.1 mg of conjugate. The starting dilution of antibody was 1:1,000 and each point on the X-axis represents a 2-fold dilution. HRP conjugated Gt-a-Rabbit IgG H&L and TMB substrate were used for detection.

ASK1-phospho-Ser83-antibody_IHC_GTX14380-1_18121409_797.jpg

GTX14380 IHC-P Image

Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ASK1 (phospho-Ser83) antibody (GTX14380)

Package List Price ($)
$ 319