Immunohistochemistry and immunoblotting were performed as previously described9 (link). The following antibodies were used: rabbit anti-fibrinogen (a gift of Dr. J. Degen), Alexa Fluor 488 isolectin GS-IB4 conjugate (#I21411, Invitrogen, Carlsbad, CA), rabbit anti-ERG (#92513, Abcam, Cambridge, MA), and mouse anti- β-actin (Sigma, St. Louis, MO).
Rabbit anti erg
Manufactured by Abcam
Sourced in United States
Rabbit anti-ERG is an antibody that recognizes the ERG protein. ERG is a transcription factor that plays a role in regulating gene expression. This antibody can be used for various applications, such as immunohistochemistry and Western blotting, to detect and analyze the presence and distribution of ERG in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Isolectin gs ib4 alexa fluor 488 conjugate, by Thermo Fisher Scientific (1 mentions)
Mouse anti β actin, by Merck Group (1 mentions)
Rabbit anti fasn, by Cell Signaling Technology (1 mentions)
Mouse anti gs, by Merck Group (1 mentions)
Fitc isolectin b4, by Vector Laboratories (1 mentions)
Rabbit anti pax6, by BioLegend (1 mentions)
Hm560, by Thermo Fisher Scientific (1 mentions)
Rabbit anti gfp, by Abcam (1 mentions)
Rat anti endomucin, by Merck Group (1 mentions)
Rabbit anti sox9, by Merck Group (1 mentions)
Rabbit anti zo 1, by Thermo Fisher Scientific (1 mentions)
Rat anti ki67, by Thermo Fisher Scientific (1 mentions)
Lsm 800, by Zeiss (1 mentions)
Rabbit anti smad3, by Cell Signaling Technology (1 mentions)
Goat igg, by Santa Cruz Biotechnology (1 mentions)
Rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Gapdh, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 488 conjugated claudin 5 monoclonal antibody, by Thermo Fisher Scientific (1 mentions)
A11006, by Thermo Fisher Scientific (1 mentions)
Plvap meca 32, by BD (1 mentions)
6 protocols using rabbit anti erg
1
Immunohistochemistry and Immunoblotting Protocols
2
Protein Expression Analysis of Myc-Cap Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lysates derived from Myc-Cap cells and appropriate tissue or cell line controls were separated by SDS/PAGE19 (link) and Western blot performed. Antibodies used were mouse anti-PSMA (clone: OTI2E1; 2 µg/mL; Thermo Fisher), rabbit anti-HPN (1 µg/mL; OriGene), mouse anti-AMACR (clone: OTI5F10; 2 µg/mL; Novus Biologicals) rabbit anti-FASN (diluted 1:1000; Cell Signaling Technology), rabbit anti-PSGR (diluted 1:500; Novus Biologicals), rabbit anti-AGR2 (clone: D9V2F; diluted 1:500; Cell Signaling Technology), rabbit anti-ERG (clone: EPR3864; diluted 1:1000; abcam), rabbit anti-CRISP3 (clone: D-6; diluted 1:100; abcam) and HRP-conjugated goat anti-rabbit and rabbit anti-mouse (diluted 1:10,000; Invitrogen).
3
Immunostaining of Cryosectioned Embryos and Flat-Mount RPE/Choroid
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cryosections, embryos were fixed in 4% paraformaldehyde (PFA), cryoprotected in 30% sucrose in phosphate-buffered saline (PBS) overnight at 4°C, embedded in OCT compound (Tissue Tec; Sakura Fine Technical, Japan), and sectioned at 10 μm using a cryostat (HM560; Thermo Fisher Scientific, Waltham, MA). Specimens were blocked with horse serum for 1 hr at room temperature and incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies for 1 hr at room temperature.
For flat-mount immunostaining of the RPE/choroid, the tissues were fixed with 4% PFA at room temperature for 30 min, and washed three times with PBS containing 0.5% Triton X-100 (PBST, Nacalai Tesque, Japan), incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies for 1 hr at room temperature (Zhu et al., 2012 (link)). The primary antibodies and dilutions used were as follows: goat anti-Aldh1a1 (1:1,000; Abcam), rabbit anti-Aldh1a3 (1:1,000; Sigma), rat anti-endomucin (1:400; Millipore), mouse anti-GS (1:1,000; Millipore), FITC-isolectin B4 (1:100; Vector Laboratories), rabbit anti-ZO-1 (1:100; Invitrogen), rabbit anti-GFP (1:1,000; Abcam), rabbit anti-ERG (1:400; Abcam), rabbit anti-Pax6 (1:200; BioLegend), and rabbit anti-Sox9 (1:200; Millipore).
For flat-mount immunostaining of the RPE/choroid, the tissues were fixed with 4% PFA at room temperature for 30 min, and washed three times with PBS containing 0.5% Triton X-100 (PBST, Nacalai Tesque, Japan), incubated with primary antibodies overnight at 4°C, followed by incubation with secondary antibodies for 1 hr at room temperature (Zhu et al., 2012 (link)). The primary antibodies and dilutions used were as follows: goat anti-Aldh1a1 (1:1,000; Abcam), rabbit anti-Aldh1a3 (1:1,000; Sigma), rat anti-endomucin (1:400; Millipore), mouse anti-GS (1:1,000; Millipore), FITC-isolectin B4 (1:100; Vector Laboratories), rabbit anti-ZO-1 (1:100; Invitrogen), rabbit anti-GFP (1:1,000; Abcam), rabbit anti-ERG (1:400; Abcam), rabbit anti-Pax6 (1:200; BioLegend), and rabbit anti-Sox9 (1:200; Millipore).
4
Quantifying Endothelial Cell Proliferation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
IHC was performed as previously described with rat anti-Ki67 (Invitrogen, Carlsbad, CA) and rabbit anti-ERG (Abcam, Cambridge, MA) as primary antibodies. ERG is a nuclear marker for endothelial cells [17 (link)] and was used to facilitate counting. Slides were examined under a confocal microscope (LSM 800, Zeiss, Jena, Germany). Analysis of endothelial cell proliferation was performed at 20X magnification (N = 5 for each group). Four random fields were sampled across the entire right lung. Endothelial cells, marked by ERG, were quantified with ImageJ and proliferating endothelial cells, marked by double staining with ERG and Ki67, were counted manually. Percent proliferating endothelial cells was calculated by dividing the number of proliferating endothelial cells by total endothelial cells.
5
Protein Interactome Analysis via IP-WB
6
Retinal Vascular Development Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To evaluate the development of retinal vessels, retinae were dissected from the eyes and fixed in 4% PFA at 4 °C overnight, followed by a rinse in phosphate-buffered saline (PBS). After being blocked in blocking buffer containing 1% fetal bovine serum with 0.25% Triton X-100 for 1 h at room temperature, the retinae were incubated overnight with Isolectin B4 (I21413, Thermo Fisher Scientific, USA), and primary antibodies including rat anti-mouse Ter-119 (553670, BD Biosciences, USA), rat anti-mouse plasmalemma vesicle-associated protein (PLVAP)/MECA-32 (553849, BD Biosciences, USA), rabbit anti-LEF1 (76010, Cell Signaling Technologies, USA), rabbit anti-ERG (ab92513, Abcam, USA), and Alexa Fluor 488-conjugated Claudin-5 monoclonal antibody (352588, Thermo Fisher Scientific, USA). The retinae were washed 3 times in PBS followed by incubation with appropriate Alexa Fluor-conjugated secondary antibodies (A-11006, A32721, or A27018, Thermo Fisher Scientific, USA) for 4 h. The Click-iT EdU Alexa Fluor-488 Imaging Kit (C10337, Thermo Fisher Scientific, USA) was used in combination with ERG antibody to detect EdU positive endothelial cells in retinae of pups that were sacrificed 3 h after intraperitoneal EdU injection.
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!