GTX14380

antibody from GeneTex

Targeting: MAP3K5 ASK1, MAPKKK5, MEKK5

Western blot (Supportive data in Antibodypedia)

ELISA (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

GTX14380

Provider

GeneTex

Proper citation

GeneTex Cat#GTX14380, RRID:AB_872826

Product name

ASK1 (phospho Ser83) antibody

Antibody type

Polyclonal

Reactivity

Human

Host

Rabbit

Western blot

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

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.

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

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

ELISA

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

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.

Immunohistochemistry

Supportive validation

Submitted by

GeneTex (provider)

Main image

Experimental details

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