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Ab4103

Manufactured by Abcam
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Sourced in United Kingdom

Ab4103 is a primary antibody designed for use in immunohistochemistry (IHC) applications. It targets a specific protein of interest. The antibody is purified and provided in a liquid format.

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Lab products found in correlation

Hpa038002, by Merck Group (2 mentions) Hpa007196, by Merck Group (2 mentions) Ab99080, by Abcam (2 mentions) Ab103328, by Abcam (2 mentions) Ab92821, by Abcam (2 mentions) Ab133836, by Abcam (2 mentions) Ab176846, by Abcam (2 mentions) Nbp1 83040, by Novus Biologicals (2 mentions) Viewplate 96 black, by PerkinElmer (2 mentions) Anti gapdh sc 47724, by Santa Cruz Biotechnology (1 mentions) Anti tubulin sc 23948, by Santa Cruz Biotechnology (1 mentions) Ab179515, by Abcam (1 mentions) Ab194297, by Abcam (1 mentions) Ab252215, by Abcam (1 mentions) Anti pro caspase 1 p10 p12, by Abcam (1 mentions) Anti β actin sc 47778, by Santa Cruz Biotechnology (1 mentions) Sc 47778, by Santa Cruz Biotechnology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Ab154343, by Abcam (1 mentions) Hoechst 33342, by Merck Group (1 mentions)

6 protocols using ab4103

1

Antibody Characterization for m6A Research

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Antibodies used in this study included a rabbit anti-m6A/m antibody (202-003, Synaptic Systems, Goettingen, Germany); a rabbit anti-human METTL3 antibody (A301-567A, Bethyl Laboratories, Inc., Montgomery, TX); a rabbit anti-human METTL14 antibody (HPA038002, Sigma, St. Louis, MO); a rabbit anti-human WTAP antibody (NBP1-83040, Novus Biologicals, LLC, Littleton, CO); a mouse anti-FTO antibody (ab92821, Abcam, Cambridge, MA); a rabbit anti-ALKBH5 antibody (HPA007196, Sigma, St. Louis, MO); a rabbit anti-human YTHDF1 antibody (ab99080, Abcam, Cambridge, MA); a rabbit anti-human YTHDF3 antibody (ab103328, Abcam, Cambridge, MA); a goat anti-mouse YTHDF3 antibody (sc-87503, Santa Cruz Inc., Dallas, TX); a rat anti-LANA antibody (ab4103, Abcam, Cambridge, MA); a rabbit anti-human YTHDF2 antibody (24744-1-AP, Proteintech Group, Rosemont, IL); a mouse anti-ORFK8 antibody (sc-57889, Santa Cruz Inc., Dallas, TX); a rabbit anti-human YTHDC1 antibody (ab133836, Abcam, Cambridge, MA); a rabbit anti-human YTHDC2 antibody (ab176846, Abcam, Cambridge, MA); a mouse anti-human β-actin antibody (sc-47778, Santa Cruz Inc., Dallas, TX). An anti-ORF57 antibody was generated by Sigma-Aldrich by immunizing a rabbit with the peptide IDGESPRFDDSIIP. The rabbit antibody to RTA and the mouse antibody to ORF65 were previously described52 (link),53 (link).
Tan B., Liu H., Zhang S., da Silva S.R., Zhang L., Meng J., Cui X., Yuan H., Sorel O., Zhang S., Huang Y, & Gao S.J. (2017). Viral and Cellular N6-Methyladenosine (m6A) and N6, 2′-O-Dimethyladenosine (m6Am) Epitranscriptomes in KSHV Life Cycle. Nature microbiology, 3(1), 108-120.
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2

Antibody Characterization for m6A Research

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Antibodies used in this study included a rabbit anti-m6A/m antibody (202-003, Synaptic Systems, Goettingen, Germany); a rabbit anti-human METTL3 antibody (A301-567A, Bethyl Laboratories, Inc., Montgomery, TX); a rabbit anti-human METTL14 antibody (HPA038002, Sigma, St. Louis, MO); a rabbit anti-human WTAP antibody (NBP1-83040, Novus Biologicals, LLC, Littleton, CO); a mouse anti-FTO antibody (ab92821, Abcam, Cambridge, MA); a rabbit anti-ALKBH5 antibody (HPA007196, Sigma, St. Louis, MO); a rabbit anti-human YTHDF1 antibody (ab99080, Abcam, Cambridge, MA); a rabbit anti-human YTHDF3 antibody (ab103328, Abcam, Cambridge, MA); a goat anti-mouse YTHDF3 antibody (sc-87503, Santa Cruz Inc., Dallas, TX); a rat anti-LANA antibody (ab4103, Abcam, Cambridge, MA); a rabbit anti-human YTHDF2 antibody (24744-1-AP, Proteintech Group, Rosemont, IL); a mouse anti-ORFK8 antibody (sc-57889, Santa Cruz Inc., Dallas, TX); a rabbit anti-human YTHDC1 antibody (ab133836, Abcam, Cambridge, MA); a rabbit anti-human YTHDC2 antibody (ab176846, Abcam, Cambridge, MA); a mouse anti-human β-actin antibody (sc-47778, Santa Cruz Inc., Dallas, TX). An anti-ORF57 antibody was generated by Sigma-Aldrich by immunizing a rabbit with the peptide IDGESPRFDDSIIP. The rabbit antibody to RTA and the mouse antibody to ORF65 were previously described52 (link),53 (link).
Tan B., Liu H., Zhang S., da Silva S.R., Zhang L., Meng J., Cui X., Yuan H., Sorel O., Zhang S., Huang Y, & Gao S.J. (2017). Viral and Cellular N6-Methyladenosine (m6A) and N6, 2′-O-Dimethyladenosine (m6Am) Epitranscriptomes in KSHV Life Cycle. Nature microbiology, 3(1), 108-120.
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3

Western Blot Analysis of Protein Expression

Cited 2 times
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Cells were lysed with RIPA lysis buffer (1 × SDS, 1 × phosphatase inhibitor cocktail, 1 × protease inhibitors) to obtain protein samples. Then, the samples were separated with SDS-PAGE and transferred to the PVDF membrane. The blots were visualized by ECL after incubating with the primary antibodies and secondary antibodies. Anti-GAPDH (sc-47724; 1:500), anti-β-actin (sc-47778; 1:500) and anti-Tubulin (sc-23948; 1:500) were purchased from Santa Cruz Biotechnology, while anti-Flag was purchased from MBL (M185-3L, Tokyo, Japan; 1:1000). Antibodies against NAT10 (ab194297; 1:1000), LANA (ab4103; 1:1000), IFI16 (ab169788; 1:1000) and ac4C (ab252215; 1:1000) were from Abcam. Anti-pro Caspase-1+p10+p12 (ab179515; 1:1000) was also obtained from Abcam. IL-1β (3A6) mouse mAb (12242; 1:1000) and cleaved Caspase-1 (Asp296) (E2G2I) rabbit mAb (89332; 1:1000) were from Cell Signaling Technology. The monoclonal rabbit anti-vIL-6 antibody (1:500) was kindly provided by Dr. Robert Yarchoan from the Center for Cancer Research, National Cancer Institute (Bethesda, Maryland, USA)53 (link),54 (link), and polyclonal rabbit anti-vIRF1 antibody (1:300) was from Dr. Gary Hayward from Viral Oncology Program, The Johns Hopkins School of Medicine (Baltimore, Maryland, USA)55 (link)–58 (link).
Yan Q., Zhou J., Wang Z., Ding X., Ma X., Li W., Jia X., Gao S.J, & Lu C. (2023). NAT10-dependent N4‐acetylcytidine modification mediates PAN RNA stability, KSHV reactivation, and IFI16-related inflammasome activation. Nature Communications, 14, 6327.
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4

Antibody-based Detection of Viral Proteins

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Primary antibodies included antibodies against METTL16 (1:1000, LS-C337738, LSBio), GAPDH (1:2000, 5174 S, Cell Signaling Technology), MAT2A (1:1000, ab154343, Abcam), RTA (1:10, a gift from Dr. Ke Lan), ORF-K8 (1:500, sc-69797, Santa Cruz), LANA (1:1000, ab4103, Abcam) and β-Actin (1:2000, sc-47778, Santa Cruz). An antibody to ORF57 was generated by immunizing the rabbits with the peptide IDGESPRFDDSIIPC. Sera collected after 5 immunizations were used at 1:4000 dilution. A monoclonal antibody to ORF65 was previously described [48 (link)] and the culture supernatant of the hybridoma was used at 1:500.
Secondary antibodies included HPR-linked goat anti-rabbit IgG (1:5000, 7074 V, Cell Signaling Technology), HPR-linked goat anti-mouse IgG (1:5000, 7076 V, Cell Signaling Technology) and HPR-linked goat anti-rat IgG (1:5000, 7077 S, Cell Signaling Technology).
Zhang X., Meng W., Feng J., Gao X., Qin C., Feng P., Huang Y, & Gao S.J. (2023). METTL16 controls Kaposi’s sarcoma-associated herpesvirus replication by regulating S-adenosylmethionine cycle. Cell Death & Disease, 14(9), 591.
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5

Quantifying KSHV Infection via Imaging

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One day prior to titration, 8,000 naïve U2OS/well were plated on viewPlate-96black (6005182, Perkin Elmer). Cells were spinoculated in the presence of 8 μg/mL of polybrene with serial dilution of precleared supernatant from infected cells. Cells were stained with antibodies against GFP (a kind gift from J. Mercer; UCL, London, United Kingdom) to detect the rKSHV.219-infected cells that express GFP from a cellular EF1α promoter or LANA (ab4103; Abcam) and Hoechst 33342.
Images from 9 fields/well were taken using Thermo Scientific Cell Insight High Content Screening System.
Gramolelli S., Elbasani E., Tuohinto K., Nurminen V., Günther T., Kallinen R.E., Kaijalainen S.P., Diaz R., Grundhoff A., Haglund C., Ziegelbauer J.M., Pellinen T., Bower M., Francois M, & Ojala P.M. (2020). Oncogenic Herpesvirus Engages Endothelial Transcription Factors SOX18 and PROX1 to Increase Viral Genome Copies and Virus Production. Cancer research, 80(15), 3116-3129.
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6

Quantifying KSHV Infection in U2OS Cells

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One day prior to titration, 8x103 naïve U2OS cells/well were plated on the viewPlate-96black (Perkin Elmer). Cells were spinoculated in the presence of 8 μg/mL of polybrene using a serial dilution of precleared supernatant from the infected LEC, BEC, ECFCLY (FIN) or ECFCBL (USA). 48h later, cells were fixed with 4% PFA, permeabilized with Triton X-100 and stained with antibodies against GFP (a kind gift from J. Mercer; UCL, London, United Kingdom) to detect the rKSHV.219-infected cells or LANA (#ab4103; Abcam) and Hoechst 33342 (Sigma). Images were taken using the automated cell imaging system ImageXpress Pico and KSHV+ cells were quantified using pipeline created in CellProfiler.
Tuohinto K., DiMaio T.A., Kiss E.A., Laakkonen P., Saharinen P., Karnezis T., Lagunoff M, & Ojala P.M. (2023). KSHV infection of endothelial precursor cells with lymphatic characteristics as a novel model for translational Kaposi’s sarcoma studies. PLOS Pathogens, 19(1), e1010753.
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