GFAP Antibody – Goat Polyclonal

Polyclonal

Goat

Human, Monkey, Rat, Bat, Bovine, Horse, Rabbit

TBS, pH 7.3, 0.02% sodium azide, 0.5% BSA

Purified from goat serum by ammonium sulphate precipitacion followed by antigen affinity chromatography using the immunizing peptide.

0.5 mg/ml (lot specific)

Human GFAP.

Aliquot and store at -20 degrees C. Avoid freeze/thaw cycles.

gene 203450

ModBase 3D Structure for P14136

GFAP

GFAP antibody was raised against Human

GFAP antibody was raised against peptide with sequence C-DGEVIKESKQEHKD, from C-Terminus of protein sequence according to NP_002046.1.

aa417-430

MBS241916 was validated for use in immunohistochemistry on a panel of 21 formalin-fixed, paraffin-embedded (FFPE) human tissues after heat induced antigen retrieval in pH 6.0 citrate buffer. After incubation with the primary antibody, slides were incubated with biotinylated secondary antibody, followed by alkaline phosphatase-streptavidin and chromogen. The stained slides were evaluated by a pathologist to confirm staining specificity. The optimal working concentration for MBS241916 was determined to be 5 ug/ml.

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 degrees 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 degrees 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