Product Page for GADD45B antibody - middle region (ARP48346_P050)

Sample Type :
Human Prostate Cancer
Primary Antibody Dilution :
2ug/mL
Color/Signal Descriptions:
GADD45B (DAB; brown), nuclei (hematoxylin; blue)
Gene Name:
GADD45B

Antibody Characterization
Antibody Catalog Number (begins with ARP) and Lot# (begins with QC#):   ARP48346_P050
Antibody Target GeneName:  GADD45B antibody
How do Aviva’s reagents play a role in your experimental goals? Antibody was used on human prostate cancer

 

Antibody Evaluation
How would you rate this antibody on a scale from 1-5 (5=best) and why?4
Would you use this antibody in future experiments? Yes
Have you used another antibody which has worked in your application? No
Do you believe the information about the reagent on Aviva’s website is correct? Yes
If the antibody works, do you plan to use it in future experiments or to publish your data? Yes
Why or why not?
How did you store the antibody after re-suspension? 4oC
Validation: IHC/ICC
Sample Description (please include species type and tissue/cell type): Human Prostate Cancer
Please explain fixation solution/method used (formalin, periodate-lysine-paraformaldehyde, acetone, etc.)?10% Neutral Buffered Formalin
How many different experimental trials were conducted using the antibody sample? Only IHC – multiple antibody concentrations and antigen retrieval conditions were tested.
Primary antibody dilution, incubation time and temperature: Primary antibody was diluted to 2ug/mL with universal antibody dilution buffer (Cat # 25886-05, Electron Microscopy Sciences, PA). The antibody incubation time was 15 min at room temperature
Secondary antibody used, dilution, incubation time and temperature: The primary antibodies were detected using Novocastra Bond Refine Polymer Detection and visualized with DAB (brown).  A hematoxylin counterstain was also applied.  The incubation time on the polymer was 8 min and DAB was 10 min.
From your IHC/ICC images, briefly explain the colors of each stain and counterstain: GADD45B (DAB; brown), nuclei (hematoxylin; blue)
Did you use an antigen retrieval method?  If so, please explain? Leica Bond antigen retrieval solution 1 (citrate buffer, pH 6.0) for 20 min at 99oC
What controls were used in your experiment? Human Prostate Cancer
Please include your detailed tissue preparation and staining procedure/protocol here:The tissue was processed into paraffin wax using standard processing procedures. Tissue sections were cut at 4um, placed on positively charged slides, and dried overnight at 37 oC before IHC staining using a Leica Bond automated immunostainer. Slides were dewaxed and heat induced antigen retrieval (HIER) was performed using citrate buffer at pH 6.0 (ER1) for 20 min.  Non specific proteins were blocked with 3% normal goat serum plus 0.1% Triton-X for 10 min and endogenous peroxidases were blocked for 10 min..  The primary antibody was diluted with universal antibody dilution buffer (Cat # 25886-05, Electron Microscopy Sciences, PA) at 2ug/mL and applied for 15 minutes at room temperature.  Primary antibodies were detected using Novocastra Bond Refine Polymer Detection and visualized with DAB (brown).  A hematoxylin counterstain was also applied.