Construction and retroviral expression of IGPR-1 in HEK-293 cells was previously described [2 (link), 4 (link)]. HA-NEDD4/pcDNA (Cat#11426), WW domain truncated NEDD4-Myc-pcMV5, GST-fusion NEDD4-WW1 (cat#104208), GST-fusion WW3 (cat#104210), and GST-fusion NEDD4-WW4 (cat#104211) all were purchased from Addgene. The development and validation of rabbit polyclonal anti-IGPR-1 antibody was previously described [1 (link)]. K48-linkage specific polyubiquitin (D9D5) Rabbit mAb (cat#8081), K63-linkage specific polyubiquitin (D7A11) Rabbit mAb (#5621), HA-Tag antibody (C29F4) Rabbit mAb (cat#3724) and Myc-Tag (71D10) Rabbit mAb (cat#2278) all were purchased from Cell Signaling Technology. Anti-ubiquitin (FK2) (cat #04-263) was purchased from Millipore-Sigma. HA-tagged wild-type ubiquitin and lysine mutant constructs were originally provided by Ted Dawson (Johns Hopkins University, Institute for Cell Engineering, Baltimore, MD) and Cam Paterson (University of North Carolina) [29 (link)]. Human NEDD4 shRNA (sc-41079-SH) which is pools of three specific 19–25 nucleotide sequences in length was purchased from Santa Cruz Biotechnology.
Myc tag 71d10 rabbit mab
Manufactured by Cell Signaling Technology
The Myc-Tag (71D10) Rabbit mAb is a monoclonal antibody that recognizes the Myc-tag, a commonly used epitope tag. The antibody can be used to detect and monitor the expression of proteins fused with the Myc-tag in various experimental applications.
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Lab products found in correlation
Hrp conjugated goat anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions)
Monoclonal anti flag hrp, by Merck Group (1 mentions)
P70 s6 kinase, by Cell Signaling Technology (1 mentions)
Wortmannin w1628, by Merck Group (1 mentions)
Hrp conjugated goat anti mouse igg h l, by Jackson ImmunoResearch (1 mentions)
Phospho 4e bp1 thr37 46, by Cell Signaling Technology (1 mentions)
2 protocols using myc tag 71d10 rabbit mab
1
IGPR-1 expression and NEDD4 interaction
2
Antibody Profiling for Autophagy Proteins
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We used the following primary antibodies: monoclonal antibodies (mAbs) against LC3 (M186-3) and ATG7 (PM039) from MBL, phospho-p70S6 kinase (#9206), p70S6 kinase (Thr389, #9202), phospho-4E-BP1 (Thr37/46, #9459), Myc-Tag (71D10) rabbit mAb (#2278 s) from Cell Signaling Technology. Rac1-102 mAb (#610650) from BD Transduction Laboratories was used to specifically detect the non-glucosylated Rac1 (Non-glu. Rac1), and Rac1 mAb 23A8 (#05-389) from Millipore was used to detect the total Rac1. Monoclonal anti-Flag-HRP (#A8592) was ordered from Sigma. The secondary antibodies HRP-conjugated goat anti-mouse IgG (H+L) (#115-035-003) and HRP-conjugated goat anti-rabbit IgG (H+L) (#111-035-003) were purchased from Jackson Immuno Research. The following chemicals were used: chloroquine (CQ), 3-Methyladenine (3-MA, M9281) and wortmannin (W1628) from Sigma, and 3-90 -2,5-diphenyltetrazolium bromide (MTT) from Amresco.
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