For immunofluorescence staining, primary antibodies were diluted in 1% PBS-BSA. Frozen sections were incubated with primary antibodies for Agrin (R&D Systems, #AF550), Hu (abcam, #96474), F4/80(BM8) (Invitrogen, #41-4801-82), Col4 (abcam, #236640), Atf4 (Invitrogen, #MA5-32364), Bax1 (SCBT, #sc-7480), Egr1 (Invitrogen, #MA5-15008), IL-18 (Invitrogen, #MA5-47203) and Hif1a (Invitrogen, #PA116601) overnight at 4 °C, followed by secondary antibodies (Alexa Fluor 647 conjugated anti-rabbit IgG; Alexa Fluor 555 conjugated anti-goat IgG; Alexa fluor 488 conjugated anti-rat; Invitrogen) for 1 h. Cell nuclei were visualized by DAPI. Sections were covered with aqueous Poly/Mount (Polyscience Inc.) and examined with a Zeiss LSM 780 laser-scanning confocal microscope (Zeiss). Images were compiled by ImageJ and Adobe Photoshop CS6 software package.
Alexa fluor 555 conjugated anti goat igg
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 555-conjugated anti-goat IgG is a fluorescently labeled secondary antibody used for the detection and visualization of goat primary antibodies in various immunoassays and imaging applications. The Alexa Fluor 555 dye provides a bright, photostable signal for sensitive detection.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions)
Alexa fluor 488 conjugated anti rabbit igg, by Thermo Fisher Scientific (3 mentions)
Sp8 confocal microscope, by Leica (2 mentions)
Fitc conjugated anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
Alexa fluor 647 conjugated anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
F4 80 bm8, by Thermo Fisher Scientific (1 mentions)
Il 18, by Thermo Fisher Scientific (1 mentions)
Hif1a, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 conjugated anti rat, by Thermo Fisher Scientific (1 mentions)
Lsm 780 laser scanning confocal microscope, by Zeiss (1 mentions)
Blocking solution, by Agilent Technologies (1 mentions)
Alexa fluor 555 conjugated anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Sc 25310, by Santa Cruz Biotechnology (1 mentions)
Sc 30820, by Santa Cruz Biotechnology (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Biosesang (1 mentions)
Anti cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Anti nrf1, by Cell Signaling Technology (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Nf κb, by Thermo Fisher Scientific (1 mentions)
6 protocols using alexa fluor 555 conjugated anti goat igg
1
Immunofluorescence Staining of Frozen Tissue Sections
2
Immunofluorescence Assay for Steroidogenic Factors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed with PBS and then fixed with 4% paraformaldehyde (Biosesang, Gyeonggi-do, Korea) for 20 min at room temperature (RT). Fixed cells were permeabilized with 0.1% Triton X-100 (Sigma) in PBS for 10 min. After permeabilization, the cells were washed twice with PBS, blocked with blocking solution (DAKO North America Inc., Carpinteria, CA, USA) for 1 h at RT, and incubated with primary antibodies overnight at 4 °C. On day 2, cells were washed twice with PBS, incubated with secondary antibodies for 1 h at RT, and then analyzed. The following antibodies were used: SF1 (1:100; LS-C162999, Lifespan Biosciences, Inc., Seattle, WA, USA), GATA4 (1:100; sc-25310, Santa Cruz Biotechnology, Santa Cruz, CA, USA); 3βHSD (1:50; sc-30820, Santa Cruz), and LHCGR (1:100; ab204950, Abcam, Cambridge, MA, USA). The secondary antibodies used were Alexa Fluor 488-conjugated anti-rabbit IgG (1:200, Invitrogen, Carlsbad, CA, USA) for SF1, Alexa Fluor 555-conjugated anti-mouse IgG (1:200, Invitrogen) for GATA4 and LHCGR, and Alexa Fluor 555-conjugated anti-goat IgG (1:100, Invitrogen) for 3βHSD.
3
Immunofluorescence Analysis of NRF1, NF-κB, and Caspase-3
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sectioned tissue was dewaxed and hydrated. Cultured cells were fixed with 4% paraformaldehyde. Samples were probed with anti-NRF1, anti-NF-κB p65, and anti-cleaved caspase-3 (9661, Cell Signaling Technology) antibodies. Binding of primary antibodies was visualized with Alexa Fluor 488-conjugated anti-rabbit IgG (R37118, Thermo) or Alexa Fluor 555-conjugated anti-goat IgG (A-21432, Thermo), and then samples were counterstained with DAPI. Images were acquired by Leica SP8 confocal microscope with 63× oil immersion objective.
4
Th1/Th2 Transcription Factors Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In order to localize the expression of Th1 cell transcription factor, T-bet, and Th2 cell transcription factor, GATA-3 immunofluorescent analysis was conducted. Only four groups were evaluated: control, OVA, dexamethasone plus OVA treatment, and 1500 mg/kg macmoondongtang plus OVA treatment. Prior to the antibody binding step, the same materials were used for immunohistochemical analysis; however, rabbit anti-mouse T-bet (Biorbyt, orb7075, Cambridge, UK) or goat anti-mouse GATA-3 (OriGene, TA305795, Rockville, MD, USA) were used as primary antibodies for 1 h at room temperature. The slides were incubated for 2 h with FITC-conjugated anti-rabbit IgG (Jackson Immunoresearch, 315-095-003, West Grove, PA, USA) or Alexa Fluor® 555-conjugated anti-goat IgG (ThermoFisher Scientific, A-21127, Waltham, MA, USA). The cells were counterstained with DAPI (ThermoFisher Scientific, 62249, Waltham, MA, USA). The images were obtained using a K1-Fluo confocal microscope (Nanoscope System, Daejeon, Korea).
5
Immunofluorescence Analysis of Immune Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In order to evaluate activation of specific proteins, such as T helper cell transcription factors (T-bet for Th1 cells and GATA-3 for Th2 cells) and NF-κB/COX-2 for inflammation pathways, immunofluorescence analyses were conducted on individuals of four groups: control, OVA treatment, OVA-induced DEX treatment, and OVA-induced 50 mg/kg KRGWE treatment. Before the antibody binding step, the same materials detailed in the immunohistochemical analyses were used, in addition to T-bet (Biorbyt, orb7075, Cambridge, UK), GATA-3 (OriGene, TA305795, Rockville, MD, USA), NF-κB (ThermoFisher Scientific), or COX-2 (Invitrogen, PA1-9032, Carlsbad, CA, USA), which were used as primary antibodies for incubation at room temperature for 1 h. All of them were incubated with FITC-conjugated anti-rabbit IgG for 2 h (#315-095-003; Jackson Immunoresearch, West Grove, PA, USA) or Alexa Fluor 555-conjugated anti-goat IgG (ThermoFisher Scientific), and cells were counterstained using 4′,6-diamidino-2-phenylindole (DAPI; ThermoFisher Scientific).
Images were produced using a K1-Fluo confocal microscope (Nanoscope System, Daejeon, South Korea), and the fluorescence intensity was measured.
Images were produced using a K1-Fluo confocal microscope (Nanoscope System, Daejeon, South Korea), and the fluorescence intensity was measured.
6
Immunofluorescence Staining of Cultured Cells and Testicular Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cultured cells were fixed with 4% paraformaldehyde for 15 min at room temperature, permeablized with triton (0.1% in TBS) for 30 min and blocked with 5% BSA in PBS for 1 h at room temperature. Cells were then incubated overnight at 4°C with anti-HIF1α (AF1935, R&D, AB_355064) and anti-STAR (8449, CST, AB_10889737). Binding of primary antibodies was visualized with Alexa Fluor 488-conjugated anti-rabbit IgG (R37118, Thermo Fisher Scientific, AB_2556546) or Alexa Fluor 555-conjugated anti-goat IgG (A-21432, Thermo Fisher Scientific, AB_2535853) for 90 min. Finally, the samples were incubated in DAPI for 10 min (Life Technologies) for nuclear counterstain. Paraffin-embedded testicular tissue was sectioned at 5 μm following dewaxing hydration treatment for immunofluorescence staining. Images were acquired using a Leica SP8 confocal microscope with 63× oil immersion objective.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!