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Our Abpromise guarantee covers the use of ab3508 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|EMSA||Use at an assay dependent concentration.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-Fr||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|WB||1/100. Detects a band of approximately 53 kDa (predicted molecular weight: 48 kDa). 1/100. Detects a band of approximately 53 kDa in COS-7 cells transfected with the human gene (predicted molecular weight: 48 kDa). This antibody supershifts DNA fragments that contain VDR response elements (e.g., rat osteocalcin and mouse osteopontin upstream elements).|
|CHIPseq||Use at an assay dependent concentration. PubMed: 21846776|
|IHC-P||1/2000 - 1/4000.|
|ChIP||Use at an assay dependent concentration. PubMed: 17244627Use at an assay dependent dilution.|
Ab3508 staining Human normal jejunum. Staining is localized to the nucleus.
Left panel: with primary antibody at 1/2000. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer, citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
ab3508 staining the Vitamin D Receptor in mVDR-transfected and untransfected Mouse NIH/3T3 cells by ICC/IF (immunocytochemistry/immunofluorescence). Cells were paraformaldehyde fixed, permeabilized with Triton X-100 and blocked with 1% BSA/5%HS for 30 minutes at 20°C. The sample was incubated with the primary antibody (1/50 in 1% BSA/5% HS in 1xPBS) for 1 hour 30 minutes at 20°C. A Cy3®-conjugated goat anti-rabbit polyclonal (1/200) was used as the secondary.
ab3508 at a 1/2000 dilution staining Vitamin D Receptor in whole Rat embryo tissue sections by Immunohistochemistry (frozen sections) incubated for 16 hours at 25°C. Samples fixed in 4% PFA prior to cutting at 30µm thickness. Blocked with 5% serum for 1 hour at 25°C. Secondary used at 1/200 polyclonal Goat anti-rabbit conjugated to Alexa Fluor 488.