SETDB1 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: SETDB1 Antibody [NBP2-20322] -

SETDB1 antibody [N2C1], Internal detects SETDB1 protein at nucleus by immunofluorescent analysis.
Sample: HeLa cells were fixed in 4% paraformaldehyde at RT for 15 min.
Green: SETDB1 stained by SETDB1 antibody [N2C1], Internal (NBP2-20322) diluted at 1:500.
Red: alpha Tubulin, a cytoskeleton marker, stained by alpha Tubulin antibody [GT114] diluted at 1:1000.
Scale bar= 10um.

Immunoprecipitation: SETDB1 Antibody [NBP2-20322] -

SETDB1 antibody [N2C1], Internal immunoprecipitates SETDB1 protein in IP experiments.
IP samples: HeLa nuclear extract
A. Control with 4 ug of preimmune Rabbit IgG
B. Immunoprecipitation of SETDB1 protein by 4 ug SETDB1 antibody [N2C1], Internal (NBP2-20322)
5 % SDS-PAGE
The immunoprecipitated SETDB1 protein was detected by SETDB1 antibody [N2C1], Internal (NBP2-20322) diluted at 1:1000.
[EasyBlot anti-rabbit IgG was used as a secondary reagent]

Chromatin Immunoprecipitation: SETDB1 Antibody [NBP2-20322] -

Cross-linked ChIP was performed with HeLa chromatin extract and 5 ug of either control rabbit IgG or anti-SETDB1 antibody. The precipitated DNA was detected by PCR with primer set targeting to p53P2.

Western Blot: SETDB1 Antibody - BSA Free [NBP2-20322] -

Demonstration of IFI16’s interaction and recruitment of specific H3K9 MTases during de novo KSHV infection.(A) TIME cells either left uninfected or infected with KSHV for 6 or 24 hr were IPed with anti-IFI16 antibodies and western blotted for the indicated proteins. (B) To confirm IFI16’s interaction with H3K9 MTases, TIME cells were infected as in (A) and IPed with antibodies against the MTases and blotted for the corresponding MTase and IFI16. (C) 293 T cells lacking IFI16 transfected with control plasmid or His-IFI16 expressing plasmid for 72 hr were utilized for His-tag pulldown using HisPur cobalt resin. Inputs and elutions were blotted for the indicated proteins. (D) 293 T cells transfected with control plasmid, IFI16 expressing plasmid or LANA expressing plasmid for 72 hr were IPed with anti-IFI16 mAb or LANA mAb. Inputs and elutions were blotted for the indicated proteins. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31682228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SETDB1 Antibody - BSA Free [NBP2-20322] -

Demonstration of IFI16’s interaction with specific H3K9 MTases in KSHV latently infected PEL (BCBL-1 and BC-3) cells and in uninfected control BJAB cells.(A) Nuclear fractions were isolated from latently infected cells and uninfected BJAB cells and treated with Benzonase. IPs were performed using anti-IFI16 mAb and LANA mAb and WBs were performed. (B) To confirm IFI16’s and LANA’s interaction with H3K9 MTases, IPs were done with Abs against the H3K9 MTases and blotted for the corresponding MTase, IFI16, LANA and HP1 alpha (heterochromatin protein 1 alpha). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31682228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SETDB1 Antibody - BSA Free [NBP2-20322] -

Demonstration of IFI16’s interaction and recruitment of specific H3K9 MTases during de novo KSHV infection.(A) TIME cells either left uninfected or infected with KSHV for 6 or 24 hr were IPed with anti-IFI16 antibodies and western blotted for the indicated proteins. (B) To confirm IFI16’s interaction with H3K9 MTases, TIME cells were infected as in (A) and IPed with antibodies against the MTases and blotted for the corresponding MTase and IFI16. (C) 293 T cells lacking IFI16 transfected with control plasmid or His-IFI16 expressing plasmid for 72 hr were utilized for His-tag pulldown using HisPur cobalt resin. Inputs and elutions were blotted for the indicated proteins. (D) 293 T cells transfected with control plasmid, IFI16 expressing plasmid or LANA expressing plasmid for 72 hr were IPed with anti-IFI16 mAb or LANA mAb. Inputs and elutions were blotted for the indicated proteins. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31682228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SETDB1 Antibody - BSA Free [NBP2-20322] -

Effect of A366 on KSHV life cycle and the demonstration of IFI16’s association with cellular H3K9 methyltransferase(s) (H3K9 MTase) and recruitment of various H3K9 MTases to the KSHV genome during de novo infection.(A) MTT cell viability assay of BCBL-1 cells treated with the H3K9me3 specific chemical inhibitor A366 at different concentrations and different time points. (B) q-RT PCR (two-step, sybr Green) of KSHV mRNAs in BCBL-1 cells treated for 72 hr with either vehicle control DMSO or A366 (10 uM and 100 uM). (C) WB of different H3 methylations and IFI16 after A366 treatment of BCBL-1 cells. (D) H3K9 methyltransferase activity (ng/h/mg) assay. TIME cells were infected with KSHV for 6 or 24 hr followed by isolation of nuclear fraction, benzonase treatment and IP with anti-IFI16 or control IgG in the presence of benzonase using the catch and release method. Elution was performed under non-denaturing conditions to keep the associated H3K9 methyltransferase active. H3K9 methyltransferase activity was assayed in the eluate (Materials and methods). *, p<0.05; **, p<0.01; ***,<0.001; unpaired t-test. (E) TIME cells were infected with KSHV genome labeled with EdU or unlabeled control KSHV (100 DNA copies/cell) for 24 hr followed by EdU-KSHV genome pulldown using Click chemistry. The inputs and eluates were blotted for different H3K9 MTases. (F) TIME cells were infected with EdU-labeled KSHV as in (D) and stained using the Click-iT EdU Alexa Fluor 594 Imaging Kit (red). Subsequently, IFA was performed against different H3K9 MTases and colocalization of the IFA signal (green) with KSHV EdU-genome staining (red) resulting in yellow was evaluated (enlarged image, white arrows). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31682228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SETDB1 Antibody - BSA Free [NBP2-20322] -

Demonstration of IFI16’s interaction with specific H3K9 MTases in KSHV latently infected PEL (BCBL-1 and BC-3) cells and in uninfected control BJAB cells.(A) Nuclear fractions were isolated from latently infected cells and uninfected BJAB cells and treated with Benzonase. IPs were performed using anti-IFI16 mAb and LANA mAb and WBs were performed. (B) To confirm IFI16’s and LANA’s interaction with H3K9 MTases, IPs were done with Abs against the H3K9 MTases and blotted for the corresponding MTase, IFI16, LANA and HP1 alpha (heterochromatin protein 1 alpha). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31682228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: SETDB1 Antibody - BSA Free [NBP2-20322] -

Demonstration of IFI16’s interaction and recruitment of specific H3K9 MTases during de novo KSHV infection.(A) TIME cells either left uninfected or infected with KSHV for 6 or 24 hr were IPed with anti-IFI16 antibodies and western blotted for the indicated proteins. (B) To confirm IFI16’s interaction with H3K9 MTases, TIME cells were infected as in (A) and IPed with antibodies against the MTases and blotted for the corresponding MTase and IFI16. (C) 293 T cells lacking IFI16 transfected with control plasmid or His-IFI16 expressing plasmid for 72 hr were utilized for His-tag pulldown using HisPur cobalt resin. Inputs and elutions were blotted for the indicated proteins. (D) 293 T cells transfected with control plasmid, IFI16 expressing plasmid or LANA expressing plasmid for 72 hr were IPed with anti-IFI16 mAb or LANA mAb. Inputs and elutions were blotted for the indicated proteins. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31682228), licensed under a CC-BY license. Not internally tested by Novus Biologicals.