AHP1017 recognises two isoforms of human High temperature requirement protein A2 (HtrA2), isoform 1 (a 45kDa protein) and isoform 2 (a 35kDa protein). Both are thought to play a role in cellular stress response as they are up-regulated following heat shock and endoplasmic reticulum stress. They promote cell death by binding and inactivating inhibitor of apoptosis proteins (IAPs), which contain baculoviral IAP repeat (BIR) binding domains. This causes an increase in caspase activity. Isoform 1 (though not 2) can also proteolytically cleave IAPs. Additionally HtrA2 isoforms can induce cell death via a mechanism dependent on its serine protease activity that is independent of IAP and caspases. Defects in HtrA2 are linked to Parkinson disease 13.
The isoforms are localised to the intermembrane space of the mitochondria to be released into the cytoplasm in response to apoptotic stimuli. Isoform 1 is ubiquitous while isoform 2 is expressed predominantly in the kidney, colon and thyroid.
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
Flow Cytometry
Immunohistology - Frozen
Immunohistology - Paraffin(1)
Immunoprecipitation
Western Blotting
0.5 - 1.0ul/ug
(1)
This product requires antigen retrieval using heat treatment prior to staining of paraffin sections. Sodium citrate buffer pH 6.0 is recommended for this purpose.
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Western Blotting
AHP1017 detects isoforms 1 and 2 of approximately 45kDa and 35kDa in U937 cell lysates
Storage
Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
Shelf Life
18 months from date of despatch.
Antiserum Preparation
Antisera to HtrA2 were raised by repeated immunisations of rabbits with highly purified antigen. Purified IgG prepared by affinity chromatography.
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