Antibody data
- Antibody Data
- Antigen structure
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- Validations
- Western blot [2]
- ELISA [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- GTX14380 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX14380, RRID:AB_872826
- Product name
- ASK1 (phospho Ser83) antibody
- Antibody type
- Polyclonal
- Reactivity
- Human
- Host
- Rabbit
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Supportive validation
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- GeneTex (provider)
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- 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.
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- GeneTex (provider)
- Main image
- Experimental details
- Western blot analysis of extracts from K562 cells using ASK1 (phospho-Ser83) antibody (GTX14380)
Supportive validation
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- GeneTex (provider)
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- 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.
Supportive validation
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- GeneTex (provider)
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- Experimental details
- Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using ASK1 (phospho-Ser83) antibody (GTX14380)