Embryos were fixed, stained, and mounted as previously described 42 . Antibodies against the following antigens were used: Dlg (mouse, 0.4 μg/ml) 43 , Arm (mouse M7A1, 0.4 μg/ml) 44 , and Slam (rabbit, 1:5,000) 45 . Secondary antibodies were labeled with Alexa dyes (Invitrogen, 0.4 μg/ml). GFP booster labeled with ATTO488 (ChromoTek, 1:500) was used for E‐Cad‐GFP.
Atto488
Manufactured by Proteintech
ATTO488 is a fluorescent dye that emits light in the green region of the visible spectrum. It is commonly used as a labeling agent for biomolecules and imaging applications.
Automatically generated - may contain errors
3 protocols using atto488
1
Immunostaining Embryonic Tissue Samples
2
Nucleolin and B23 Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nucleolin was detected with a rabbit polyclonal antibody raised against purified human nucleolin (pab0971-P Covalabs / dil. WB 1/1000) as previously described [15 (link), 19 (link)]. B23 was detected with a mouse monoclonal antibody (FC82291 Sigma-Aldrich #B0556 / dil. WB 1/1000). α-tubulin was detected with a mouse monoclonal antibody (DM1A Sigma-Aldrich #T9026 / dil. WB 1/1000). β-actin was detected with a mouse monoclonal antibody (AC-15 Sigma-Aldrich #A5441 / dil. WB 1/1000). Acetylated tubulin was detected with a mouse monoclonal antibody (611B1 Sigma-Aldrich #T7451 / dil. IF 1/300). GFP detection was boosted with an anti GFP antibody directly coupled to Atto488 (Chromotek / dil. IF 1/200). For Immunofluorescence, secondary antibodies were coupled to Alexa555 (Molecular Probes DaMAlexa555 #A31570 / dil. 1/2000). For WB, secondary antibodies were coupled to IRdye800 (Li-Cor #92632211 / dil. 1/2500) or Alexa680 (Li-Cor #92632220 / dil. 1/15000). EB3-tagRFP plasmid has been previously described [15 (link)].
3
Multicolor Labeling of Synaptic Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasmids were kindly provided by our collaborators: pCMV–GFP–GPI and pCMV–HA-Nlg1 (Olivier Thoumine, Bordeaux/Peter Scheiffele, Basel), pCMV–Homer–GFP (Daniel Choquet, Bordeaux), pSyn5-HA-neurexin1ß (Markus Missler, Münster).
Primary antibodies used were the following: anti-GFP mouse IgG, monoclonal antibody (200 μg; clones 7.1 and 13.1; Catalogue number 11814460001; Roche Diagnostics) and anti-HA high affinity, rat monoclonal antibody (clone 3F10; 50 μg; Catalogue number 11867423001; Roche Diagnostics), anti-GFP nanobodies ATTO488 (Chromotek).
Secondary antibodies used were the following: Qdot655 goat F(ab′)2 anti-mouse IgG conjugate (H+L) (1 μM solution Q11022MP Lot729518; Invitrogen/Molecular Probes). For dual colour imaging the following were used: Qdot605 goat F(ab')2 anti-mouse IgG conjugate (H+L) (1 μM solution, Q11002MP; Invitrogen/Molecular Probes) plus Qdot655 goat F(ab')2 anti-rat IgG conjugate (H+L)(1 μM solution, Q11621MP; Invitrogen/Molecular Probes).
Primary antibodies used were the following: anti-GFP mouse IgG, monoclonal antibody (200 μg; clones 7.1 and 13.1; Catalogue number 11814460001; Roche Diagnostics) and anti-HA high affinity, rat monoclonal antibody (clone 3F10; 50 μg; Catalogue number 11867423001; Roche Diagnostics), anti-GFP nanobodies ATTO488 (Chromotek).
Secondary antibodies used were the following: Qdot655 goat F(ab′)2 anti-mouse IgG conjugate (H+L) (1 μM solution Q11022MP Lot729518; Invitrogen/Molecular Probes). For dual colour imaging the following were used: Qdot605 goat F(ab')2 anti-mouse IgG conjugate (H+L) (1 μM solution, Q11002MP; Invitrogen/Molecular Probes) plus Qdot655 goat F(ab')2 anti-rat IgG conjugate (H+L)(1 μM solution, Q11621MP; Invitrogen/Molecular Probes).
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!