HSP90AA Antibody – Mouse Monoclonal

Monoclonal

IgG

2G5.G3

Mouse

Human, Mouse, Rat

PBS, pH 7.2; 50% glycerol, 0.09% azide

Protein G Purified from tissue culture supernatant

1 mg/ml (lot specific)

Hsp90 alpha specific (>96% alpha specific by ELISA).

Short term 4 degree C, long term aliquot and store at -20 degree C, avoid freeze thaw cycles.

HSP90AA / HSP90 Alpha

Purified protein

HSP90AA / HSP90 Alpha antibody was raised against human Hsp90alpha

Due to the highly specific nature of antibodies and antigens, we cannot predict or be held responsible with respect to how this antibody will behave in your systems. Researchers using this antibody should conduct optimization studies to achieve the most optimal result possible for their intended application.

The following protocol is a recommendation only, and MyBioSource, Inc. makes no guarantee of the results:

Tissue Preparation:
Formalin fixation and embedding in paraffin wax.

Tissue Sectioning:
Make 4-um sections and place on pre-cleaned and charged microscope slides. Heat in a tissue-dryingoven for 45 minutes at 60°C.

Deparaffinization:
Wash dry slides in 3 changes of xylene - 5 minutes each @ RT

Rehydration:
Wash slides in 3 changes of 100% alcohol - 3 minutes each @ RT
Wash slides in 2 changes of 95% alcohol - 3 minutes each @ RT
Wash slides in 1 change of 80% alcohol - 3 minutes @ RT
Rinse slides in gentle running distilled water - 5 minutes @ RT

Antigen retrieval:
Steam slides in 0.01 M sodium citrate buffer, pH 6.0 at 99-100°C - 20 minutes
Remove from heat and let stand at room temperature in buffer - 20 minutes
Rinse in 1X TBS with Tween (TBST) -1 minute @ RT

Immunostaining:
(Do not allow tissues to dry at any time during the staining procedure)
Apply a universal protein block - 20 minutes @ RT
Drain protein block from slides, apply diluted primary antibody - 45 minutes @ RT
Rinse slides in 1 X TBST - 1 minute @ RT
Apply a biotinylated secondary antibody appropriate for the primary antibody - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase streptavidin - 30 minutes @ RT
Rinse slides in 1X TBST -1 minute @ RT
Apply alkaline phosphatase chromogen substrate - 30 minutes @ RT
Wash slides in distilled water - 1 minute @ RT

Dehydrate:
(This method should only be used if the chromogen substrate is alcohol insoluble (e.g. Vector Red, DAB)
Wash slides in 2 changes of 80% alcohol - 1 minute each @ RT
Wash slides in 2 changes of 95% alcohol - 1 minute each @ RT
Wash slides in 3 changes of 100% alcohol - 1 minute each @ RT
Wash slides in 3 changes of xylene - 1 minute each @ RT
Apply coverslip