Octamer-4 (Oct-4), a member of the POU family of transcription factors, has been demonstrated to be vital for the formation of self-renewing pluripotent stem cells.
During embryogenesis, expression of Oct-4 is limited to pluripotent cells of the inner cell mass (ICM) that contribute to the formation of all fetal cell types.
Applications:Suitable for use in Western Blot, Immunohistochemistry and Immunocytochemistry.
Other applications not tested.
Recommended Dilutions:Western Blot: 1:200-1:1000 using ECLImmunohistochemistry: 1:200-1:1000Immunocytochemistry: 1:200-1:1000Optimal dilutions to be determined by the researcher.
Immunohistochemistry Protocol:1.
Dissect tissues and freeze on dry ice
2.
Cut on a cryostat, 10um slices
3.
Dry slides at RT for 30-90 min
4.
Fix with cold Acetone:Methanol (50:50) 2 min.
Either process further or store at -20ºC.
Alternative is fixation with 4% PFA and treatment with 0.05% trypsin solution (incubation 10 min.
at 37ºC, followed by 3 washes with PBS)
5.
Let air dry
6.
PBS 5 min
7.
Incubate in 50mM ammonium chloride 30 min
8.
PBS 5 min
9.
Blocking serum 30-45 min
10.
Primary antibody (O5403) 90 min
11.
Wash 3X for 5 min
12.
Secondary fluorescent conjug.
antibody 30 min
13.
Wash 3X for 5 min
14.
Mount.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：100ug
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by affinity chromatography
Form：Supplied as a liquid in PBS, pH 7.4. No preservative added.
Specificity：Recognizes human Oct-4 (POU5F1) at ~38kD. Species Crossreactivity: rodent.
Isotype：IgG
Calc Applications Abbrev：IC IHC WB
Calc Crossreactivity：Hu Rd
Immunogen：Synthetic peptide corresponding to aa282-297 of human Oct-4.
