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Anti gw182

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Anti-GW182 is a laboratory product that targets the GW182 protein. GW182 is a key component of the RNA-induced silencing complex (RISC), which plays a crucial role in gene silencing and RNA interference (RNAi) pathways. The Anti-GW182 product is intended for use in research applications involving the study of GW182 and its functions in these biological processes.

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Lab products found in correlation

Aj1236b, by Thermo Fisher Scientific (6 mentions) Anti gapdh, by Merck Group (3 mentions) Anti actin, by Merck Group (2 mentions) Anti myc, by Cell Signaling Technology (2 mentions) Anti puromycin, by Merck Group (2 mentions) Stabilized peroxidase conjugated secondary antibodies, by Thermo Fisher Scientific (2 mentions) Anti flag, by Merck Group (2 mentions) Anti ha, by Merck Group (2 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions) Anti gfp, by Proteintech (2 mentions) Anti flag, by Proteintech (2 mentions) Anti e2f1, by Abcam (2 mentions) Anti ago1, by Abcam (2 mentions) Anti ago2, by Abnova (2 mentions) Anti dapk1, by Abcam (2 mentions) Anti dapk3, by Abcepta (2 mentions) Anti tnrc6b, by Abnova (2 mentions) Anti tnrc6c, by Fortis Life Sciences (2 mentions) Anti ago3, by Proteintech (2 mentions) Anti cd99, by Proteintech (2 mentions)

7 protocols using anti gw182

1

Characterization of Ago1x Antibody Specificity

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Polyclonal anti‐Ago1x antibody was generated in rabbit against the unique peptide sequence, RQNAVTSLDRRKLSKP, and was affinity‐purified (Pierce Biotechnology). To confirm the specificity, the anti‐Ago1x antibody was incubated with the same peptide at a final concentration of 1 μg/ml for 2 h with gentle agitation at room temperature before using it for Western blot (described below). Anti‐Ago1 antibody (Cell Signaling Technologies, 9388; Sigma, SAB4200065), anti‐Dicer (Sigma, SAB4200087), anti‐HA (Sigma, clone 3F10, 11867423001; and Cell Signaling Technology, clone 6E2, 2367), anti‐GW182 (Bethyl Laboratories, A302‐329A), anti‐puromycin (Merck Millipore, clone 12D10, MABE343), anti‐Myc (Cell Signaling Technologies, 22725), anti‐FLAG (Sigma, clone M2, F1804), anti‐Actin (Sigma, A3854), anti‐GAPDH (Sigma, G9295), and stabilized peroxidase‐conjugated secondary antibodies (Thermo Fisher Scientific) were used at the manufacturer‐recommended dilutions. Alexa Fluor‐conjugated secondary antibodies (Molecular Probes) were used for immunofluorescence.
Singh A., Manjunath L.E., Kundu P., Sahoo S., Das A., Suma H.R., Fox P.L, & Eswarappa S.M. (2019). Let‐7a‐regulated translational readthrough of mammalian AGO1 generates a microRNA pathway inhibitor. The EMBO Journal, 38(16), e100727.
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2

Antibody Validation for mRNA Decay Factors

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Antibodies used in this study were as follows: rabbit polyclonal anti-DDX6 (Bethyl Laboratories), rabbit polyclonal anti-Pat1b (Bethyl Laboratories), rabbit polyclonal anti-DCP2 (Bethyl Laboratories), mouse monoclonal anti-EDC3 (Abcam), rabbit polyclonal anti-GW182 (Bethyl Laboratories), mouse monoclonal anti-eIF4E (BD Transduction Laboratories), rabbit polyclonal anti-PABP (Abcam), rabbit monoclonal anti-Ago2 (Cell Signaling), mouse monoclonal anti-myc (BioShop Canada Inc.), mouse monoclonal anti-β-actin (Sigma). Rabbit polyclonal antibodies against CNOT1 and CNOT7 were kindly provided by T. Yamamoto.
Rouya C., Siddiqui N., Morita M., Duchaine T.F., Fabian M.R, & Sonenberg N. (2014). Human DDX6 effects miRNA-mediated gene silencing via direct binding to CNOT1. RNA, 20(9), 1398-1409.
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3

Characterization of DAPK and miRNA pathway

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HeLa and 293Tcells were cultured in MEDM plus 10% FBS. Both cells were determined to be free from mycoplasma contamination. The antibodies used were from different vendors: Anti-DAPK1 (Abcam, ab109382; ABclonal, A5741), Anti-DAPK2 (Abgent, AP7033A), Anti-DAPK3 (Abgent, AJ1236b; Thermal scientific, PA5–27700), Anti-ACTIN (PTGCN, 60008-I-Ig), Anti-E2F1 (Abcam, ab179445), Anti-GW182 (Bethyl, A302–329A), Anti-TNRC6B (Abnova), Anti-TNRC6C (Bethyl, A303–969A), Anti-Ago1 (Abcam, ab105104), Anti-Ago2 (Abnova, H00027161-M01), Anti-Ago3 (Proteintech, 19692-I-AP), Anti-Ago4 (CST, 6913S), Anti-CD99 (Proteintech, 60354-I-Ig; ABclonal, A2028), Anti-NDRG1 (Abcam, ab124698), Anti-SAFB (Abnova, H00006294-M04), Anti-FLAG (ABclonal, AE005), Anti-GFP (Proteintech, 66002-I-Ig), anti-Mcherry (ABclonal, AE002). Western blotting results were quantified by the ImageJ software.
Zhang K., Zhang X., Cai Z., Zhou J., Cao R., Zhao Y., Chen Z., Wang D., Ruan W., Zhao Q., Liu G., Xue Y., Qin Y., Zhou B., Wu L., Nilsen T., Zhou Y, & Fu X.D. (2018). A Novel Class of MicroRNA Recognition Elements That Function Only in Open Reading Frames. Nature structural & molecular biology, 25(11), 1019-1027.
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4

Characterization of DAPK and miRNA pathway

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HeLa and 293Tcells were cultured in MEDM plus 10% FBS. Both cells were determined to be free from mycoplasma contamination. The antibodies used were from different vendors: Anti-DAPK1 (Abcam, ab109382; ABclonal, A5741), Anti-DAPK2 (Abgent, AP7033A), Anti-DAPK3 (Abgent, AJ1236b; Thermal scientific, PA5–27700), Anti-ACTIN (PTGCN, 60008-I-Ig), Anti-E2F1 (Abcam, ab179445), Anti-GW182 (Bethyl, A302–329A), Anti-TNRC6B (Abnova), Anti-TNRC6C (Bethyl, A303–969A), Anti-Ago1 (Abcam, ab105104), Anti-Ago2 (Abnova, H00027161-M01), Anti-Ago3 (Proteintech, 19692-I-AP), Anti-Ago4 (CST, 6913S), Anti-CD99 (Proteintech, 60354-I-Ig; ABclonal, A2028), Anti-NDRG1 (Abcam, ab124698), Anti-SAFB (Abnova, H00006294-M04), Anti-FLAG (ABclonal, AE005), Anti-GFP (Proteintech, 66002-I-Ig), anti-Mcherry (ABclonal, AE002). Western blotting results were quantified by the ImageJ software.
Zhang K., Zhang X., Cai Z., Zhou J., Cao R., Zhao Y., Chen Z., Wang D., Ruan W., Zhao Q., Liu G., Xue Y., Qin Y., Zhou B., Wu L., Nilsen T., Zhou Y, & Fu X.D. (2018). A Novel Class of MicroRNA Recognition Elements That Function Only in Open Reading Frames. Nature structural & molecular biology, 25(11), 1019-1027.
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5

Characterization of Anti-Ago1x Antibody

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Polyclonal anti-Ago1x antibody was generated in rabbit against the unique peptide sequence, RQNAVTSLDRRKLSKP, and was affinity-purified (Pierce Biotechnology). To confirm the specificity, the anti-Ago1x antibody was incubated with the same peptide at a final concentration of 1 μg/ml for 2 h with gentle agitation at room temperature before using it for Western blot (described below). Anti-Ago1 antibody (Cell Signaling Technologies, 9388; Sigma, SAB4200065), anti-Dicer (Sigma, SAB4200087), anti-HA (Sigma, clone 3F10, 11867423001; and Cell Signaling Technology, clone 6E2, 2367), anti-GW182 (Bethyl Laboratories, A302-329A), anti-puromycin (Merck Millipore, clone 12D10, MABE343), anti-Myc (Cell Signaling Technologies, 22725), anti-FLAG (Sigma, clone M2, F1804), anti-Actin (Sigma, A3854), anti-GAPDH (Sigma, G9295), and stabilized peroxidase-conjugated secondary antibodies (Thermo Fisher Scientific) were used at the manufacturer-recommended dilutions. Alexa Fluor-conjugated secondary antibodies (Molecular Probes) were used for immunofluorescence.
Singh A., Manjunath L.E., Kundu P., Sahoo S., Das A., Suma H.R., Fox P.L, & Eswarappa S.M. (2019). Let‐7a‐regulated translational readthrough of mammalian AGO1 generates a microRNA pathway inhibitor. The EMBO Journal, 38(16), e100727.
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6

Protein Detection and Quantification

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Western blotting was performed using the Novex NuPAGE SDS/PAGE gel system (Invitrogen). Total cell lysates were run either on 3–8% Tris-acetate or 4–12% Bis-Tris precast gels, transferred to nitrocellulose membranes, and probed with antibodies specific to proteins of interest. Detection and quantification of blots were performed on Amersham hyperfilm ECL (Cytiva #28906839) and developed on film processor SRX-101A (Konica). Antibodies used for western blots were obtained from commercial sources as follows: anti-GW182 (Bethyl #A302-239A), anti-Ago2 (Cell Signaling #2897), anti-PABP1 (Cell Signaling #4992), anti-RPL26 (Bethyl #A300-686A), anti-GAPDH (Sigma #G8795), anti-β-actin (Sigma #A2228), anti-GFP (Roche #11814460001), anti-tubulin (Sigma-Aldrich #T9026), anti-HA (Cell Signaling #C29F4), anti-rabbit IgG, HRP-conjugated (GE Healthcare #NA934), and anti-mouse IgG, HRP-conjugated (GE Healthcare #NA931).
La Rocca G., King B., Shui B., Li X., Zhang M., Akat K.M., Ogrodowski P., Mastroleo C., Chen K., Cavalieri V., Ma Y., Anelli V., Betel D., Vidigal J., Tuschl T., Meister G., Thompson C.B., Lindsten T., Haigis K, & Ventura A. (2021). Inducible and reversible inhibition of miRNA-mediated gene repression in vivo. eLife, 10, e70948.
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7

Antibody Validation for CNOT Complex

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Antibodies against CNOT1 and CNOT3 have been previously described (Chen et al., 2011) .
Commercially available antibodies used in this study were: anti-a-tubulin (Sigma, T9026), anti-PARP (CST, 9542S), anti-CNOT9 (Proteintech, 22503-1-AP), anti-Flag (MBL, PM020), anti-Xrn-1(Bethyl A300-443A), anti-HistoneH3 (CST, 4499S), anti-GW182 (Bethyl, A302-329A), anti-CNOT2 (CST, 34214S), anti-4EBP1 (CST, 9644S), anti-b-actin (CST, 4970L), anti-GAPDH (CST, 2118L), anti-CNOT10 (Bethyl A304-899A), and anti-Ago2 (CST, 2897S). small colonies were picked and seeded into individual wells of a 48 well plate and propagated further. Knockout clones were confirmed by DNA sequencing and Western blotting.
Sarmah H., Ito K., Kaneko M., Abe T., & Yamamoto T. (2020). Loss of CNOT9 begets impairment in gastrulation leading to embryonic lethality.
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