Anti-GDNF Receptor alpha 1/GFRA1 Antibody Picoband®

Western blot analysis of GFRA1 using anti-GFRA1 antibody (PB9202).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours.
Lane 1: recombinant human GFRA1 protein 0.5 ng.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-GFRA1 antigen affinity purified polyclonal antibody (Catalog # PB9202) 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 GFRA1 at approximately 39 kDa. The expected band size for GFRA1 is at 39 kDa.

Western blot analysis of GFRA1 using anti-GFRA1 antibody (PB9202).
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 30 ug of sample under reducing conditions.
Lane 1: human placenta tissue lysates,
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-GFRA1 antigen affinity purified polyclonal antibody (Catalog # PB9202) 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 GFRA1 at approximately 51 kDa. The expected band size for GFRA1 is at 51 kDa.

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

Relative expression of specific genes in spermatogonial cells. (A) The mRNA expression of TET1 in spermatogonial cells was detected by QRT-PCR. (B) The mRNA expression of PLZF in spermatogonial cells was detected by QRT-PCR. (C) The mRNA expression of GFRα1 in spermatogonial cells was detected by QRT-PCR. (D) The mRNA expression of PRDM1 in spermatogonial cells was detected by QRT-PCR. (E) The mRNA expression of MAGE4 in spermatogonial cells was detected by QRT-PCR. (F) The expression of PLZF in TET1 overexpressed cells was detected by Western Blot. (G) Quantification of PLZF protein levels in TET1 overexpressed cells. (H) The expression of GFRα1 in TET1 overexpressed cells was detected by Western Blot. (I) Quantification of GFRα1 protein levels in TET1 overexpressed cells. p < .0001(****), p < .001(***), p < .01(**), p < .05(*).
Index in PubMed under a CC BY license. PMID: 35222097

IHC analysis of GFRA1 using anti-GFRA1 antibody (PB9202).
GFRA1 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-GFRA1 Antibody (PB9202) 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.