Full length protein, corresponding to amino acids 1-410 of Human RGL4
General notesFor use in immunofluorescence, antibody must be purified:
1. Wash and equilibrate rProtein A Sepharose with 5 bed volumes of Protein A IgG Binding Buffer (Thermo Scientific), 3 times.
2. Load 50 µL sample containing IgG of interest into the micro-centrifuge tube containing 50 µL rProtein A sepharose, and add 200 µL of binding buffer, total volume 300 µL.
3. The sample and rProtein A sepharose were incubated on a 3D rocking platform at the room temperature for 2hrs.
4. Wash and equilibrate the spin column (Micro Bio-Spin Chromatography Column, Bio-Rad Laboratories) with IgG binding buffer.
5. Add the sample mixture from step 3 to the spin column.
6. Centrifuge at 2000 rpm for 5 seconds to collect flow through.
7. Wash column with 300 µL IgG binding buffer 3 times to remove unbound and non-specifically bound proteins.
8. Elute the bound IgG in 100 µL fractions into micro-centrifuge tubes containing 10 µL 1M Tris, pH 10 respectively. Collect at least 6 fractions.
9. Detect purified protein concentration by OD 280. Collect fractions containing purified
10. Dialyze purified protein against PBS at 4oC
11. Detect protein concentration by OD 280.
12. Store the purified IgG at -20oC
Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesICC/IF: Use at a concentration of 10 µg/ml. Must be purified before use.
WB: 1/500 - 1/1000. Detects a band of approximately 52 kDa (predicted molecular weight: 52 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Immunohistochemical analysis of HeLa cells labelled with purified ab67650 at 10 µg/ml followed by fluorescein-conjugated secondary antibody.
Cells were fixed in 4% paraformaldehyde and permeabilised with 0.1% Triton X-100.
References for Anti-RGL4 antibody (ab67650)
has not yet been referenced specifically in any publications.
Publishing research using ab67650? Please let us know so that we can cite the reference in this datasheet.
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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"