The DNA sequence encoding a part of zebrafish Pou5f3 (residues 1-122) was amplified by PCR and ligated into the pET21 vector to produce a histidine (His)-tagged protein. The recombinant protein was expressed in E. coli and purified by SDS-PAGE, followed by electroelution in Tris-glycine buffer without SDS. The purified protein was dialyzed against 1 mM HEPES (pH 7.5), lyophilized, and used for injection into two mice and one rabbit. The obtained antisera were affinity-purified with recombinant Pou5f3-His protein electroblotted onto a membrane (Immobilon; EMD Millipore, IPVH00010). Similarly, a part of Pabpc1l (residues 6-348) was expressed in E. coli, purified, and used for injection into two mice. The obtained antisera were affinity-purified with recombinant Pabpc1l protein electroblotted onto the membrane. The crude extracts from oocytes and embryos at appropriate time points were separated by SDS-PAGE, blotted onto the Immobilon membrane, and probed with the anti-Pou5f3 mouse and rabbit antibodies and with the anti-Pabpc1l mouse antibody. Rpl11 was detected by immunoblotting the crude extracts with anti-Rpl11 rabbit antibody (Abcam; ab79352).
Ab79352
Manufactured by Abcam
Ab79352 is a laboratory equipment product. It serves a core function related to scientific research and analysis. No further details are available regarding its specific intended use or application.
Automatically generated - may contain errors
Lab products found in correlation
Immobilon, by Merck Group (1 mentions)
Mouse anti rps6, by Cell Signaling Technology (1 mentions)
Mouse anti armadillo n2 7a1, by Developmental Studies Hybridoma Bank (1 mentions)
Rabbit anti fibrillarin, by Abcam (1 mentions)
Mouse anti fibrillarin, by Abcam (1 mentions)
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
2 protocols using ab79352
1
Zebrafish Pou5f3 and Pabpc1l Protein Purification and Antibody Generation
2
Immunohistochemistry of Drosophila Imaginal Discs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Eye-antennal imaginal discs and salivary glands were prepared for immunohistochemistry using standard protocols. As the growth conditions strongly affect salivary gland size, all the experiments were controlled by synchronization of L3 wandering larvae after a single-day egg collection. To further control this issue, a controlled the egg laying for 5 h was set up, and the salivary glands were analysed 96 h after egg laying (96–101 h AEL; electronic supplementary material, figure S4g–j′).
Primary antibodies used were: mouse anti-Armadillo N27A1 at 1 : 100 (Developmental Studies Hybridoma Bank, DSHB), mouse anti-Dlg at 1 : 1000 (4F3, DSHB), rabbit anti-Viriato (Vito) at 1 : 250 (ABGent), rat anti-DCad at 1 : 100, mouse anti-AH6 at 1 : 10 (DSHB), rabbit anti-Fibrillarin at 1 : 250 (Abcam, #ab5821), mouse anti-Fibrillarin at 1 : 500 (Abcam, #ab4566), mouse anti-RpS6 at 1 : 100 (Cell Signaling, #2317), mouse anti-RpL11 at 1 : 100 (Abcam, #ab79352), mouse anti-RpL10A at 1 : 400 (Abcam, #ab55544), rabbit anti-RpL22 at 1 : 100 (kind gift from Dr Vassie Ware). To stain for cellular limits phalloidin conjugated with rhodamine was used at a dilution of 1 : 1000. Appropriate Alexa-Fluor conjugated secondary antibodies were from Molecular Probes. Images were obtained with the Leica SP2 confocal system or Leica SP5 confocal system and processed with Adobe Photoshop .
Primary antibodies used were: mouse anti-Armadillo N27A1 at 1 : 100 (Developmental Studies Hybridoma Bank, DSHB), mouse anti-Dlg at 1 : 1000 (4F3, DSHB), rabbit anti-Viriato (Vito) at 1 : 250 (ABGent), rat anti-DCad at 1 : 100, mouse anti-AH6 at 1 : 10 (DSHB), rabbit anti-Fibrillarin at 1 : 250 (Abcam, #ab5821), mouse anti-Fibrillarin at 1 : 500 (Abcam, #ab4566), mouse anti-RpS6 at 1 : 100 (Cell Signaling, #2317), mouse anti-RpL11 at 1 : 100 (Abcam, #ab79352), mouse anti-RpL10A at 1 : 400 (Abcam, #ab55544), rabbit anti-RpL22 at 1 : 100 (kind gift from Dr Vassie Ware). To stain for cellular limits phalloidin conjugated with rhodamine was used at a dilution of 1 : 1000. Appropriate Alexa-Fluor conjugated secondary antibodies were from Molecular Probes. Images were obtained with the Leica SP2 confocal system or Leica SP5 confocal system and processed with Adobe P
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!