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Ab184224

Manufactured by Abcam
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Ab184224 is an antibody product manufactured by Abcam. It is a primary antibody intended for use in various research applications. The core function of this product is to specifically bind to and detect its target antigen.

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Lab products found in correlation

Ab15568, by Abcam (2 mentions) Novex 10 20 tricine protein gels, by Thermo Fisher Scientific (2 mentions) α tubulin, by Merck Group (2 mentions) Alexa fluor fluorescent secondary antibody, by Thermo Fisher Scientific (2 mentions) α strep tag, by Abcam (2 mentions) α flag tag, by Merck Group (2 mentions) α wnv capsid, by Abcam (2 mentions) α wnv capsid, by GeneTex (2 mentions) α wnv capsid, by Merck Group (2 mentions) α denv capsid, by GeneTex (2 mentions) α zikv capsid, by GeneTex (2 mentions) α fibrillarin, by Thermo Fisher Scientific (2 mentions) α fibrillarin, by Cell Signaling Technology (2 mentions) α ddx55, by Fortis Life Sciences (2 mentions) α pym1, by Novus Biologicals (2 mentions) α magoh, by Santa Cruz Biotechnology (2 mentions) α rbm8a, by Merck Group (2 mentions) α aif, by Cell Signaling Technology (2 mentions) α lamin b1, by Abcam (2 mentions) Hrp conjugated secondary antibodies α mouse or α rabbit, by Cytiva (2 mentions)

7 protocols using ab184224

1

Immunoblotting Analysis of Viral Proteins

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Lysates from the above mentioned assays were analyzed by SDS-PAGE, using standard 12% SDS-PAGE to resolve mCherry and its XP-fusion variants, and precast Novex™ 10–20% tricine protein gels (Thermo Fisher) to resolve XPs and enterovirus 2B. Proteins were then transferred to 0.2 µm nitrocellulose membranes and blocked with 4% Marvel milk powder in phosphate-buffered saline (PBS). Immunoblotting of mCherry was performed using anti-mCherry antibody (Abcam, ab167453, 1:3000). A custom rabbit polyclonal antibody raised against XP peptide SNSGNRVSQDQNLQ (GenScript; only able to detect strongly overexpressed XP, 1:250) and an anti-Strep mouse antibody (Abcam, ab184224, 1:1000) were used for detecting HAstV1 XP and Strep-tagged proteins, respectively. The following antibodies were used for cellular targets: anti-tubulin (Abcam, ab15568, 1:500), anti-VDAC1 (Abcam, ab14734, 1:1000), and anti-LAMIN A + C (Abcam, ab133256, 1:3000). Immunoblots were imaged on a LI-COR ODYSSEY CLx imager and analyzed using Image Studio™ version 5.2.
Lulla V, & Firth A.E. (2020). A hidden gene in astroviruses encodes a viroporin. Nature Communications, 11, 4070.
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2

Immunoblot Analysis of Enterovirus Proteins

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Lysates from virus-infected or pCAG-transfected cells were analyzed by SDS-PAGE, using standard 12% SDS-PAGE to resolve virus structural proteins and precast Novex™ 10–20% tricine protein gels (Thermo fisher) to resolve UP. Proteins were then transferred to 0.2 µm nitrocellulose membranes and blocked with 4% Marvel milk powder in phosphate-buffered saline (PBS). Immunoblotting of the enterovirus VP3 structural protein was performed using Enterovirus pan monoclonal antibody (Thermo Fisher, MA5-18206) at 1:1,000 dilution. A custom rabbit polyclonal antibody raised against C-terminal UP peptide CPPRKPEPMRLG (GenScript), an anti-Strep mouse antibody (Abcam, ab184224), and an anti-HA mouse antibody (Abcam, ab130275) were used for detection of EV7 UP, EV7 Strep-tagged UP, and PV1 HA-tagged UP, respectively. The following antibodies were used for cellular targets: anti-tubulin (Abcam, ab15568), anti-VDAC1 (Abcam, ab14734), anti-GAPDH (Ambio, AM4300) and anti-calnexin (Merck, MAB3126). To ensure synchronicity of infection, a high MOI was used for virus infections. Immunoblots were imaged and analyzed on a LI-COR imager. The original LICOR scans and quantifications are shown in Fig. S11.
Lulla V., Dinan A.M., Hosmillo M., Chaudhry Y., Sherry L., Irigoyen N., Nayak K.M., Stonehouse N.J., Zilbauer M., Goodfellow I, & Firth A.E. (2018). An Upstream Protein-Coding Region in Enteroviruses Modulates Virus Infection in Gut Epithelial Cells. Nature microbiology, 4(2), 280-292.
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3

Antibody Panel for Flavivirus Detection

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The following antibodies were used: α-strep tag (ab184224, abcam), α-FLAG tag (F7425, Sigma), α-V5 (A190–120A, Bethyl), α-tubulin (T6199, Sigma), α-WNV capsid (abcam, ab21673), α-WNV capsid (Genetex, GTX131947 ), α-WNV capsid (Sigma, SAB3500912), α-DENV capsid (Genetex, GTX103343), α-ZIKV capsid (Genetex, GTX133317), α-fibrillarin (Thermo Fisher Scientific, AMSA6771), α-fibrillarin (Cell Signaling Technologies, 2639S), α-DDX55 (Bethyl, A303–027A), α-PYM1 (Novus, NBP2–46366), α-MAGOH (Santa Cruz, sc-56724), α-RBM8A (Sigma, HPA018403), α-AIF (Cell Signaling, 5318S), α-Lamin B1 (Abcam, ab231282). The α-flavivirus glycoprotein (4G2) hybridoma was provided by Dr. M. Diamond (Washington University). Alexa-Fluor fluorescent secondary antibodies were from Life Technologies. HRP-conjugated secondary antibodies (α-mouse or α-rabbit) were from Amersham.
Li M., Johnson J.R., Truong B., Kim G., Weinbren N., Dittmar M., Shah P.S., Von Dollen J., Newton B.W., Jang G.M., Krogan N.J., Cherry S, & Ramage H. (2019). Identification of Antiviral Roles for the Exon-Junction Complex and Nonsense-Mediated Decay in Flaviviral Infection. Nature microbiology, 4(6), 985-995.
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4

Antibody Panel for Flavivirus Detection

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The following antibodies were used: α-strep tag (ab184224, abcam), α-FLAG tag (F7425, Sigma), α-V5 (A190–120A, Bethyl), α-tubulin (T6199, Sigma), α-WNV capsid (abcam, ab21673), α-WNV capsid (Genetex, GTX131947 ), α-WNV capsid (Sigma, SAB3500912), α-DENV capsid (Genetex, GTX103343), α-ZIKV capsid (Genetex, GTX133317), α-fibrillarin (Thermo Fisher Scientific, AMSA6771), α-fibrillarin (Cell Signaling Technologies, 2639S), α-DDX55 (Bethyl, A303–027A), α-PYM1 (Novus, NBP2–46366), α-MAGOH (Santa Cruz, sc-56724), α-RBM8A (Sigma, HPA018403), α-AIF (Cell Signaling, 5318S), α-Lamin B1 (Abcam, ab231282). The α-flavivirus glycoprotein (4G2) hybridoma was provided by Dr. M. Diamond (Washington University). Alexa-Fluor fluorescent secondary antibodies were from Life Technologies. HRP-conjugated secondary antibodies (α-mouse or α-rabbit) were from Amersham.
Li M., Johnson J.R., Truong B., Kim G., Weinbren N., Dittmar M., Shah P.S., Von Dollen J., Newton B.W., Jang G.M., Krogan N.J., Cherry S, & Ramage H. (2019). Identification of Antiviral Roles for the Exon-Junction Complex and Nonsense-Mediated Decay in Flaviviral Infection. Nature microbiology, 4(6), 985-995.
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5

Western Blot Analysis of Membrane Proteins

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Cell lysates were prepared in 4x loading buffer 200 mM Tris, pH 6.8, 8% SDS, 40% glycerol and 0.02% bromophenol blue under reducing conditions (+ dithiothreitol) and heated to 80°C for 10 min. Samples were separated by SDS-PAGE and transferred to a nitrocellulose membrane using the Trans-Blot Turbo Transfer System (Bio-Rad). Membranes were blocked in 5% powdered milk in PBS-T (0.05% Tween-20 in PBS) for 1 h at room temperature with constant agitation. Membranes were washed with PBS-T and incubated with primary antibody anti-CD147 (ab232967, Abcam), anti-ASCT2 (SLC1A5) (V501, Cell Signaling), anti-CD98 (12206-T62, sino biologicals), anti-E1, anti-Strep (ab184224 or ab180957, Abcam) and anti-E2 (NR44002, Bei Resources) overnight at 4°C. After washing with PBS-T, the membranes were incubated the secondary antibody anti-mouse (ab6728, Abcam) or anti-rabbit (ab6721, Abcam) for 1 h at room temperature. Detection was done with the Amersham ECL Western Blotting Detection Reagent (GE healthcare).
De Caluwé L., Coppens S., Vereecken K., Daled S., Dhaenens M., Van Ostade X., Deforce D., Ariën K.K, & Bartholomeeusen K. (2021). The CD147 Protein Complex Is Involved in Entry of Chikungunya Virus and Related Alphaviruses in Human Cells. Frontiers in Microbiology, 12, 615165.
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6

Overexpression of Ostreococcus CNNM Genes

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Ostreococcus cultures were grown for 6–7 days in artificial sea water (ASW) under 12/12 h light/dark cycles at 20 °C, under enriched blue light (183 Moonlight Blue Filter), as previously described [37 (link)]. New transgenic lines overexpressing OtCNNM1 (ostta05g01490) and OtCNNM2 (ostta11g01030) genes were PCR-amplified using specific primers incorporating the STREP tag sequence (Table S2). Then, fragments were digested with AvrII (NEB) and ligated into the pOTOX vector [22 (link)]. Genomic transformation was conducted as previously published [38 (link)] and performed under the CCA1-LUC background line [22 (link)]. Positive colonies were selected with ClonNAT antibiotic (Nourseothricin, Jena Bioscience) and protein samples were validated by immunoblotting using an anti-STREP antibody (Strep-II antibody, abcam ab184224).
Gil S., Feord H.K, & van Ooijen G. (2023). Homologs of Ancestral CNNM Proteins Affect Magnesium Homeostasis and Circadian Rhythmicity in a Model Eukaryotic Cell. International Journal of Molecular Sciences, 24(3), 2273.
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7

Multimerization of overexpressed XP

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To analyze multimerization of overexpressed XP, electroporation of Huh7.5.1 cells was performed in full media at 220 V and 975 µF using a Bio-Rad Gene Pulser. At 20 hpe, cells were washed with PBS and co-immunoprecipitation was performed using the Pierce™ Magnetic HA-Tag IP/Co-IP Kit (Thermo Scientific) according to the manufacturer’s instructions. Aliquots of input (5%) and immunoprecipitation fractions were analyzed by western blot using the indicated tag-specific antibodies: anti-HA (Abcam, ab20084, 1:1000) and anti-Strep (Abcam, ab184224, 1:1000).
Lulla V, & Firth A.E. (2020). A hidden gene in astroviruses encodes a viroporin. Nature Communications, 11, 4070.
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