goat polyclonal anti gfra1  (R&D Systems)

Journal: bioRxiv

Article Title: Hedgehog-dependent and Hedgehog-independent roles for Growth Arrest Specific 1 in mammalian kidney morphogenesis

doi: 10.1101/2024.05.02.592197

Figure Lengend Snippet: RET receptors are expressed normally in Gas1 -/- metanephros. (A-H) Whole-mount in situ hybridization for Ret (A-D) and Gfra1 (E-H) in wildtype (A,C,E,G) and Gas1 -/- (B,D,F,H) metanephros (MetN). (I-P) Immunofluorescent antibody detection of RET (I-L) and GFRA1 (M-P) in coronal kidney sections from E11.5 and E12.0 wildtype (I,K,M,O) and Gas1 mutant (J,L,N,P) embryos. Asterisks denote Gas1 mutant kidneys that have not undergone ureteric bud (UB) branching morphogenesis (insets; L,P). (Q) Quantitation of Etv5 MM fluorescence intensity normalized to MM area (3 sections/kidney) in E11.5 wildtype (n=6 kidneys) and Gas1 -/- (n=5 kidneys) embryos. Scale bars (A,C,E,G) 100μm; (I) 50μm. P-values were determined by a two-tailed Student’s t-test: non-significant: p>0.05 and significant: p≤0.05.

Article Snippet: Sections were blocked (1X PBS+ 10% Donkey serum + 0.3% Triton) for 1 h at 37°C, then incubated with rabbit anti-RET (1:200, Cell Signaling, Cat#3223) and goat anti-GAS1 (1:900, R&D Systems; Cat#AF2544) or goat anti- GFRA1 (1:500, R&D Systems, Cat# AF560) overnight at 4°C.

Techniques: In Situ Hybridization, Mutagenesis, Quantitation Assay, Fluorescence, Two Tailed Test

Journal: bioRxiv

Article Title: Hedgehog-dependent and Hedgehog-independent roles for Growth Arrest Specific 1 in mammalian kidney morphogenesis

doi: 10.1101/2024.05.02.592197

Figure Lengend Snippet: GAS1 is secreted from the MM and accumulates in the UB epithelium. (A-J) Immunofluorescent detection of DAPI (gray; A,F), GAS1 (cyan; B,G), RET (magenta; C,H), HSPG2 (yellow; D,I) in metanephros coronal sections from E12.0 wildtype (A-E) and Gas1 -/- (F-J) embryos. GAS1/RET/HSPG2 merged images are shown (E,J). Dotted lines circle the ureteric bud (UB; B,G,C,H). (K) Western blot detection of immunoprecipitated endogenous GAS1 from supernatants collected from cultured wildtype and Gas1 mutant metanephric mesenchyme (MM) cells from E18.5 kidneys (n=3 technical replicates). (L-Q) Representative images of Proximity Ligation Assay (PLA) detection of GFRA1/RET PLA+ signal (yellow; M,P) merged with CDH1 (cyan; N,Q) antibody detection in metanephros coronal sections from E11.5 wildtype (L-N) and Gas1 -/- (O-Q) embryos. DAPI labels nuclei (gray; L,O). Dotted lines mark the UB (M,P). (R) Quantitation of the total number of GFRA1/RET PLA+ puncta (normalized to CDH1+ cells; 3 sections/kidney) in E11.5 wildtype (n=11) and Gas1 -/- (n=4) metanephros. (S-X) Representative images of PLA detection of GAS1/RET PLA+ signal (yellow; T,W) merged with CDH1 (cyan; U,X) antibody detection in metanephros coronal sections from E11.5 wildtype (S-U) and Gas1 -/- (V-X) embryos. DAPI labels nuclei (gray; S,V). Dotted lines mark the UB (T,W). (Y) Quantitation of the total number of GAS1/RET PLA+ puncta (normalized to CDH1+ cells; 3 sections/kidney) in E11.5 wildtype (n=16) and Gas1 -/- (n=7) metanephros. (Z-C’) Whole-mount in situ hybridization of Wnt11 expression in wildtype (Z,A’) and Gas1 -/- (B,C’) kidney explants cultured for 4 days in the presence or absence of recombinant murine GDNF (75ng/mL). Dotted lines denote the metanephros (Z’B’) or UB (B’; inset). (D’) Quantitation of the total number of Wnt11 -expressing UB branch tips in non-treated and GDNF-treated control (wildtype or Gas1 +/- ; n=8 non-treated, n=12 GDNF-treated) and Gas1 -/- (n=7 non-treated, n=8 treated) kidney explant cultures. Scale bar (F,N) 25μm; (Z) 250μm. P-values were determined by a two-tailed Student’s t-test (R,Y) or an ordinary one-way ANOVA with Tukey’s multiple comparisons test (D’): p>0.05 and significant: p≤0.05.

Article Snippet: Sections were blocked (1X PBS+ 10% Donkey serum + 0.3% Triton) for 1 h at 37°C, then incubated with rabbit anti-RET (1:200, Cell Signaling, Cat#3223) and goat anti-GAS1 (1:900, R&D Systems; Cat#AF2544) or goat anti- GFRA1 (1:500, R&D Systems, Cat# AF560) overnight at 4°C.

Techniques: Western Blot, Immunoprecipitation, Cell Culture, Mutagenesis, Proximity Ligation Assay, Quantitation Assay, In Situ Hybridization, Expressing, Recombinant, Control, Two Tailed Test

Journal: Stem Cell Research & Therapy

Article Title: Expression profiling of stemness markers in testicular germline stem cells from neonatal and adult Swiss albino mice during their transdifferentiation in vitro

doi: 10.1186/s13287-024-03701-8

Figure Lengend Snippet: Sequences of primers used in the real time and RT-PCR experiment

Article Snippet: Primary antibodies (GFRA1, ITGA6, ALPL, CD9, NES, TBXT and PDX-1), goat anti-rabbit IgG-FITC, goat anti-rabbit IgG- Alexa Fluor 488, Rabbit anti-goat IgG- Alexa Fluor 488, (Santa Cruz Biotechnology, CA, USA); CD9 antibody (BioLegend, San Diego, CA, USA); Reverse Transcriptase PCR primers (Sigma Genosys, Bangalore, India); Taq DNA polymerase, 100 bp & 1 kb ladder (New England Biolabs, MA, USA); DAPI, TRI reagent, Ethidium Bromide, agarose, glycine, Trizma Base and paraformaldehyde were purchased from Sigma- Aldrich, MO, USA.

Techniques: Derivative Assay

Journal: Stem Cell Research & Therapy

Article Title: Expression profiling of stemness markers in testicular germline stem cells from neonatal and adult Swiss albino mice during their transdifferentiation in vitro

doi: 10.1186/s13287-024-03701-8

Figure Lengend Snippet: Confocal microscopy of SSC colonies derived from neonatal mice testis in day 24 of culture, showing the expression of GFRA1 ( A ), ITGA6 ( B ) and ALPL ( C ) and CD9 ( D ). The corresponding phase contrast images ( E - H ) of the SSC colonies are also shown. Bar = 20 μm. Real time PCR analysis of the expression of stem cell markers and a germ cell marker in SSC colonies on day 24 of culture ( I ). Bar = 20 μm

Article Snippet: Primary antibodies (GFRA1, ITGA6, ALPL, CD9, NES, TBXT and PDX-1), goat anti-rabbit IgG-FITC, goat anti-rabbit IgG- Alexa Fluor 488, Rabbit anti-goat IgG- Alexa Fluor 488, (Santa Cruz Biotechnology, CA, USA); CD9 antibody (BioLegend, San Diego, CA, USA); Reverse Transcriptase PCR primers (Sigma Genosys, Bangalore, India); Taq DNA polymerase, 100 bp & 1 kb ladder (New England Biolabs, MA, USA); DAPI, TRI reagent, Ethidium Bromide, agarose, glycine, Trizma Base and paraformaldehyde were purchased from Sigma- Aldrich, MO, USA.

Techniques: Confocal Microscopy, Derivative Assay, Expressing, Real-time Polymerase Chain Reaction, Marker

Journal: Stem Cell Research & Therapy

Article Title: Expression profiling of stemness markers in testicular germline stem cells from neonatal and adult Swiss albino mice during their transdifferentiation in vitro

doi: 10.1186/s13287-024-03701-8

Figure Lengend Snippet: Expression of GFRA1 ( A ), ITGA6 ( B ), ALPL ( C ) and CD9 ( D ) in dispersed cells of SSCs derived from adult mice testicular cells. The corresponding DAPI-stained images are shown in E - H . Bar = 20 μm

Article Snippet: Primary antibodies (GFRA1, ITGA6, ALPL, CD9, NES, TBXT and PDX-1), goat anti-rabbit IgG-FITC, goat anti-rabbit IgG- Alexa Fluor 488, Rabbit anti-goat IgG- Alexa Fluor 488, (Santa Cruz Biotechnology, CA, USA); CD9 antibody (BioLegend, San Diego, CA, USA); Reverse Transcriptase PCR primers (Sigma Genosys, Bangalore, India); Taq DNA polymerase, 100 bp & 1 kb ladder (New England Biolabs, MA, USA); DAPI, TRI reagent, Ethidium Bromide, agarose, glycine, Trizma Base and paraformaldehyde were purchased from Sigma- Aldrich, MO, USA.

Techniques: Expressing, Derivative Assay, Staining