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Immunoprecipitation and Western Blot using Aldolase polyclonal antibody (Product # PA1-27236). 300 µl aliquots of whole anti-aldolase antiserum were used to precipitate varying amounts of purified aldolase. Precipitates with controls were compared by SDS-PAGE and Western blot. Samples shown in the image are: 1. Purified aldolase 2. 300 µl antiserum with no antigen (negative control) 3. 300 µl antiserum with ~100 µl aldolase (2.5 mg/ml) 4. 300 µl antiserum with ~200 µl aldolase (2.5 mg/ml). For the precipitation, 300 ul of antiserum and an equal volume of aldolase antigen in PBS was incubated for 24 hrs at 4°C, centrifuged for 6 minutes at 13K RPM, washed once with PBS, centrifuged and dissolved in 60 ul 0.1 N NaOH. 90 ul of PBS was added, the sample was divided into 2 portions, and an equal volume of reducing (+4% BME) or non-reducing 2X sample buffer was added. The reduced samples were boiled for five minutes, and all samples were run at 140 V for ~45 minutes on a 4-20% tris/glycine gradient gel. Gel was stained, destained and imaged using standard protocols. Precipitation of aldolase was confirmed by comparison of increasing amounts of antigen with the control protein by SDS PAGE and observation of a 40-45 kD MW band corresponding to Aldolase. Additional higher and lower molecular weight bands correspond to serum proteins.
|Tested species reactivity||Rabbit|
|Host / Isotype||Goat / IgG|
|Immunogen||Full length protein from rabbit muscle aldolase.|
|Storage buffer||0.02M potassium phosphate, pH 7.2, with 0.15M NaCl, 10mg/ml BSA|
|Contains||0.01% gentamicin sulfate|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-27236 detects Aldolase from rabbit samples.
PA1-27236 has been successfully used in ELISA and Western blot procedures.
The PA1-27236 immunogen is full length protein from rabbit muscle aldolase.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.