BD, 611203, BD Transduction Laboratories™ Purified Mouse Anti-Oct3/4

Alternative Name: Oct3, OTF3, Oct4, OTF4, POU5F1
Reactivity: Mouse (QC Testing), Human (Tested in Development)
Isotype: Mouse IgG1, κ
Immunogen: Mouse Oct3 aa. 252-372 Recombinant Protein
Application: Bioimaging, Western blot (Routinely Tested), Immunofluorescence (Tested During Development)
Target Molecular Weight: 46 kDa
Concentration: 250 µg/ml
RRID: AB_398736
Storage Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at -20°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Recommended Assay Procedures: Recommended Assay Procedures Bioimaging 1. Seed the cells in appropriate culture medium at an appropriate cell density in a BD Falcon™ 96-well Imaging Plate (Cat. No. 353219), and culture overnight to 48 hours. 2. Remove the culture medium from the wells, wash the wells twice with 100 µl of 1x PBS, and fix the cells by adding 100 µl of fresh 3.7% Formaldehyde in PBS or BD Cytofix™ fixation buffer (Cat. No. 554655) to each well and incubating for 10 minutes at room temperature (RT). 3. Remove the fixative from the wells, and wash the wells twice with 100 µl of 1x PBS. 4. Permeabilize the cells by adding 100 µl of 1× BD Perm/Wash™ buffer (Cat. No. 554723) to each well and incubating for 30 minutes at RT. 5. Remove the permeabilizer, and wash the wells twice with 100 µl of 1x PBS. 6. Dilute the antibody in BD Perm/Wash™ buffer, and stain the cells by adding 50 µl of the diluted antibody to each well and incubating for 1 hour at RT. 7. Remove the diluted antibody, and wash the wells three times with 100 µl of 1x PBS. 8. Remove the PBS, dilute the second-step reagent in BD Perm/Wash™ buffer, and stain the cells by adding 50 µl of the diluted second-step reagent to each well and incubating for 1 hour at RT. 9. Remove the diluted second-step reagent, and wash the wells twice with 100 µl of 1x PBS. 10. Remove the PBS, and counter-stain the nuclei by adding 100 µl of a 2 µg/ml solution of Hoechst 33342 (eg, Sigma-Aldrich Cat. No. B2261) in 1x PBS to each well at least 15 minutes before imaging. 11. View and analyze the cells on an appropriate imaging instrument.