Anti-DAZL Picoband® Antibody

Western blot analysis of DAZL using anti-DAZL antibody (A02069-2).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
Lane 2: rat testicular tissue lysates,
Lane 3: mouse testicular tissue lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DAZL antigen affinity purified polyclonal antibody (Catalog # A02069-2) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for DAZL at approximately 38KD. The expected band size for DAZL is at 33KD.

Effects of EZH2 interference or overexpression and JMJD3 interference or overexpression on self-renewal, proliferation and differentiation of spermatogonia. ( A ) The mRNA levels of PCNA, Cyclin-A, GFRA1, PLZF and C-KIT related to spermatogonia self-renewal and proliferation were changed after EZH2 and JMJD3 knockdown. ( B ) The expression of PCNA, cyclin-A, GFRA1, PLZF, C-KIT, DAZL and VASA was detected by qRT-PCR after EZH2 and JMJD3 overexpression. ( C ) The protein expression changes as well as statistical analysis of PCNA, DAZL and GFRA1 after EZH2 and JMJD3 overexpression. The membrane is lysed prior to hybridization with the antibody and the image has been cropped for a more aesthetically pleasing display. The full- length blots can be obtained from Additional file 2: Fig . ( D ) The cell cycle of EZH2 and JMJD3 overexpression cells was detected by flow cytometry. ( E ) Protein interaction network of EZH2, JMJD3 and spermatogonia self-renewal, proliferation and differentiation-related genes
Index in PubMed under a CC BY license. PMID: 38424516

IHC analysis of DAZL using anti-DAZL antibody (A02069-2).
DAZL was detected in paraffin-embedded section of human testies cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DAZL Antibody (A02069-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of DAZL using anti-DAZL antibody (A02069-2).
DAZL was detected in paraffin-embedded section of mouse testies cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DAZL Antibody (A02069-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of DAZL using anti-DAZL antibody (A02069-2).
DAZL was detected in paraffin-embedded section of rat testies cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-DAZL Antibody (A02069-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

Flow Cytometry analysis of HEPA1-6 cells using anti-DAZL antibody (A02069-2).
Overlay histogram showing HEPA1-6 cells stained with A02069-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DAZL Antibody (A02069-2, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of HL-60 cells using anti-DAZL antibody (A02069-2).
Overlay histogram showing HL-60 cells stained with A02069-2 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-DAZL Antibody (A02069-2, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.