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Product name
Phospho S300 PDH E1 alpha protein (PDHA1) ELISA Kit
See all Pyruvate Dehydrogenase E1-alpha subunit products (4) ...
Precision
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| 1 | 8 | 2.5% |
| Sample | n | Mean | SD | CV% |
|---|---|---|---|---|
| 1 | 4 | 7.8% |
Sample type
Cell culture extracts, Tissue Extracts
Assay type
Quantitative
Sensitivity
15 µg/ml
Range
15 µg/ml - 500 µg/ml
Cross reactivity
Reacts with
Mouse, Rat, Cow, Human
Product overview
ab115345 is an in vitro enzyme-linked immunosorbent assay to determine the levels of phospho S300 PDHA1 protein in cell and tissue lysates. The assay employs a mouse antibody specific for PDHA1 protein coated on a 96-well plate. Samples are pipetted into the wells and PDHA1 protein present in the sample is bound to the wells by the immobilized antibody. The wells are washed and a rabbit anti-phospho S300 PDHA1 protein detector antibody is added. After washing away unbound detector antibody, HRP-conjugated anti-rabbit antibody is pipetted into the wells. The wells are again washed, an HRP substrate solution (TMB) is added to the wells and color develops in proportion to the amount of phospho S300 PDHA1 protein bound. The developing blue color is measured at 600 nm. Optionally the reaction can be stopped by adding hydrochloric acid which changes the color from blue to yellow and the intensity can be measured at 450 nm.
Notes
Store all components at 4°C. This kit is stable for 6 months from
receipt. Unused microplate strips should be returned to the pouch
containing the desiccant and resealed.
receipt. Unused microplate strips should be returned to the pouch
containing the desiccant and resealed.
Tested applications
ELISAmore details
Properties
Storage instructions
Please see Notes section
| Components | Identifier | 1 x 96 tests |
|---|---|---|
| 10X Blocking Buffer | P/N 8209802 | 1 x 6ml |
| 10X HRP Label | 1 x 1ml | |
| 10X phospho S300 PDHA1 protein Detector Antibody | 1 x 0.7ml | |
| 20x Buffer | P/N 8203013 | 1 x 20ml |
| Extraction Buffer | P/N 8201093 | 1 x 15ml |
| Microplate strips | 1 unit | |
| TMB Development Solution | P/N 8209803 | 1 x 12ml |
Research areas
Metabolism >> Pathways and Processes >> Metabolic signaling pathways >> Energy transfer pathways >> Energy Metabolism
Metabolism >> Pathways and Processes >> Mitochondrial Metabolism >> Mitochondrial markers
Kits/ Lysates/ Other >> Kits >> Cell Metabolism Kits >> Dehydrogenase Kits
Signal Transduction >> Metabolism >> Mitochondrial
Signal Transduction >> Metabolism >> Energy Metabolism
Alternative names
- MS967
Applications
Show applications key
Our Abpromise guarantee covers the use of ab115345 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA
Phospho S300 PDH E1 alpha protein (PDHA1) ELISA Kit images:
ELISA - Phospho S300 PDH E1 alpha protein (PDHA1) ELISA Kit (ab115345)
The phosphorylation state of PDHA1 can vary by treatment but also by cell culture conditions such as media supplements, nutrients and also cell density.
- Phospho S300 PDH E1 alpha protein (PDHA1) ELISA Kit (ab115345)
HeLa cells were cultured for 4 hours in media supplemented with DCA (20mM) to specifically inhibit mitochondrial PDH kinases, or NaF (20mM), a general inhibitor of serine/threonine protein phosphatases. The DCA treatment did not reduce the level of phospho S300 confirming that there is little endogenous phospho S300. Conversely NaF treatment, to inhibit cellular serine phosphatases, did increase the phosphorylation level of S300.
- Phospho S300 PDH E1 alpha protein (PDHA1) ELISA Kit (ab115345)
The PDHA1 bound from undosed HeLa cells was subject to in-well kinase treatment (PDK1&3) or in-well phosphatase treatment(PDP1) according to the supplementary protocol shown below. Untreated cells did not show a significant endogenous phosphorylation signal at S300 this could be increased by kinase treatment. Phosphatase treatement had no effect on the endogenous (low) level of phospho S300 signal.
Protocols
References for Phospho S300 PDH E1 alpha protein (PDHA1) ELISA Kit (ab115345)
ab115345 has not yet been referenced specifically in any publications.
Publishing research using ab115345? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"