The following primary antibodies were used: mouse anti-Wingless (1:1000 for standard protocol and 1:300 for extracellular staining; DSHB 4D4), rabbit anti-HA (1:1000, Cell Signalling), rat anti-OLLAS (1:10, Abnova), chicken anti-GFP (1:1000, Abcam), mouse anti-Integrin (1:100, DSHB CF.6G11), mouse anti-Dlp (1:50, DSHB 13G8), guinea pig anti-Senseless (1:1000, gift from H. Bellen), mouse anti-V5 (1:500, Invitrogen), goat anti-GMAP (1:100, gift from S. Munro, MRC-LMB Cambridge) guinea pig anti-Godzilla (1:1500; 22 (link)). Secondary antibodies used were Alexa 488 and Alexa 555 (1:500 for standard protocol and 1:250 for extracellular staining, Molecular Probes). Total and extracellular immunostaining of imaginal discs was performed as previously described 4 (link). Imaginal discs were mounted in Vectashield with DAPI (Vector Laboratories) and imaged using either a Leica SP5 or LSM710 confocal microscope. Z stacks were generated with 1-1.5 micron intervals. Confocal images were processed with ImageJ (N.I.H), Volocity (PerkinElmer), ZEN2.0 (Zeiss) and Photoshop CS5.1 (Adobe). All XY confocal images show a single confocal section, XZ and YZ projections were created using Volocity. Adult wings were mounted in Euparal (Fisher Scientific) and imaged with a Zeiss Axiophot2 microscope with an Axiocam HRC camera.
Rat anti ollas
Manufactured by Abnova
The Rat anti-OLLAS is a monoclonal antibody that specifically recognizes the OLLAS tag, a synthetic peptide epitope. This antibody can be used for the detection and purification of recombinant proteins containing the OLLAS tag.
Automatically generated - may contain errors
Lab products found in correlation
Chicken anti gfp, by Abcam (3 mentions)
Axiocam hrc camera, by Zeiss (2 mentions)
Alexa 488, by Thermo Fisher Scientific (2 mentions)
Lsm710, by Leica (2 mentions)
Axiophot 2 microscope, by Zeiss (2 mentions)
Mouse anti v5, by Thermo Fisher Scientific (2 mentions)
Rabbit anti ha, by Cell Signaling Technology (2 mentions)
Zen 2, by Zeiss (2 mentions)
Alexa 555, by Thermo Fisher Scientific (2 mentions)
Vectashield with dapi, by Vector Laboratories (2 mentions)
Euparal, by Thermo Fisher Scientific (2 mentions)
Volocity, by PerkinElmer (2 mentions)
Lsm800 confocal microscope, by Zeiss (1 mentions)
Alexa fluor conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Axiocam 503 camera, by Zeiss (1 mentions)
Rabbit anti gfp, by Abcam (1 mentions)
Chicken anti β galactosidase, by Abcam (1 mentions)
Zen 2 blue edition imaging software, by Zeiss (1 mentions)
3 protocols using rat anti ollas
1
Comprehensive Immunostaining Protocol
2
Comprehensive Immunostaining Protocol
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The following primary antibodies were used: mouse anti-Wingless (1:1000 for standard protocol and 1:300 for extracellular staining; DSHB 4D4), rabbit anti-HA (1:1000, Cell Signalling), rat anti-OLLAS (1:10, Abnova), chicken anti-GFP (1:1000, Abcam), mouse anti-Integrin (1:100, DSHB CF.6G11), mouse anti-Dlp (1:50, DSHB 13G8), guinea pig anti-Senseless (1:1000, gift from H. Bellen), mouse anti-V5 (1:500, Invitrogen), goat anti-GMAP (1:100, gift from S. Munro, MRC-LMB Cambridge) guinea pig anti-Godzilla (1:1500; 22 (link)). Secondary antibodies used were Alexa 488 and Alexa 555 (1:500 for standard protocol and 1:250 for extracellular staining, Molecular Probes). Total and extracellular immunostaining of imaginal discs was performed as previously described 4 (link). Imaginal discs were mounted in Vectashield with DAPI (Vector Laboratories) and imaged using either a Leica SP5 or LSM710 confocal microscope. Z stacks were generated with 1-1.5 micron intervals. Confocal images were processed with ImageJ (N.I.H), Volocity (PerkinElmer), ZEN2.0 (Zeiss) and Photoshop CS5.1 (Adobe). All XY confocal images show a single confocal section, XZ and YZ projections were created using Volocity. Adult wings were mounted in Euparal (Fisher Scientific) and imaged with a Zeiss Axiophot2 microscope with an Axiocam HRC camera.
3
Embryonic Development Antibody Staining
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Embryos were fixated and stained as described (Müller, 2008) . Primary antibodies used were: guinea pig anti-Alk (1:1000 (Loren et al., 2003) ), rabbit anti-Alk (1:750 (Loren et al., 2003) ), chicken anti-β-galactosidase (1:200; Abcam ab9361), mouse anti-Fasciclin3 (1:50; DSHB 7G10), mouse anti-Antp (1:50; DSHB 4C3) rabbit anti-GFP (1:500; Abcam ab290), chicken anti-GFP (1:300; Abcam ab13970), mouse anti-Wg (1:50, DSHB 4D4), rat anti-OLLAS (1:200, preabsorbed on w 1118 embryos; Abnova), rabbit anti-Org-1 (1:1000, (Mendoza-Garcia et al., 2017)), sheep anti-digoxygenin-AP fab fragment 1:4000 (Roche). Alexa Fluor®-conjugated secondary antibodies were from Jackson Immuno Research. Embryos were dehydrated in an ascending ethanol series before clearing and mounting in methylsalicylate. Images were acquired with a Zeiss LSM800 confocal microscope or Axiocam 503 camera, processed and analyzed employing Zeiss ZEN2 (Blue Edition) imaging software. Nuclei quantification (Fig. 6G) was performed with ImageJ software (Schneider et al., 2012) . Raw images were converted into binary format and nuclei were quantified with 3D nuclei counter package (Bolte and Cordelieres, 2006) .
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