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H6908

Manufactured by Abcam
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H6908 is a laboratory equipment product. It is a piece of laboratory equipment that serves a core function. No further details about its intended use or application can be provided in an unbiased and factual manner.

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Lab products found in correlation

Pierce ecl, by Thermo Fisher Scientific (3 mentions) Ab8580, by Merck Group (1 mentions) Ab85392, by Abcam (1 mentions) Ab4729, by Abcam (1 mentions) Formaldehyde, by Merck Group (1 mentions) Ab9106, by Abcam (1 mentions) S32354, by Thermo Fisher Scientific (1 mentions) Alexa fluor 568 phalloidin, by Thermo Fisher Scientific (1 mentions) Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions) Ab3114, by Santa Cruz Biotechnology (1 mentions) Odyssey software, by LI COR (1 mentions) Amersham protran membrane, by GE Healthcare (1 mentions) P irf3, by Abcam (1 mentions) Secondary abs, by LI COR (1 mentions) Odyssey infrared imaging system, by LI COR (1 mentions) F7425, by Merck Group (1 mentions) α tubulin, by Merck Group (1 mentions) Ab1162, by Abcam (1 mentions) Ab31940, by Abcam (1 mentions) Ab167453, by Merck Group (1 mentions)

8 protocols using h6908

1

ChIP-Seq Profiling of Histone Marks

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Standard ChIP assays were performed as described in Supplemental Experimental Procedures with antibodies against H3K27me3 (Millipore, #07-449), H3K4me3 (Abcam, #ab8580), HA tag (Sigma, #H6908), Jmjd3 (Abcam, #ab85392) or normal rabbit IgG.
Pan D., Huang L., Zhu L.J., Zou T., Ou J., Zhou W, & Wang Y.X. (2015). Jmjd3-mediated H3K27me3 dynamics orchestrate brown fat development and regulate white fat plasticity. Developmental cell, 35(5), 568-583.
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2

ChIP-seq and ChIP-qPCR of HA-T-VP64

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mCherry+ 416B cells were sorted by FACS (24 h after dox addition), then expanded without dox before HA-T-VP64 expression was induced for 48 h and cells crosslinked using 1% formaldehyde (Sigma). ChIP-qPCR and ChIP-seq were performed and analysed essentially as described previously (Wilson et al., 2010b (link)) using anti-HA (Sigma, H6908) and anti-H3K27Ac (Abcam, ab4729) antibodies, and the ChIP-qPCR primers listed in supplementary material Table S6. ChIP-seq data are available at GEO with accession number GSE61189.
Wilkinson A.C., Kawata V.K., Schütte J., Gao X., Antoniou S., Baumann C., Woodhouse S., Hannah R., Tanaka Y., Swiers G., Moignard V., Fisher J., Hidetoshi S., Tijssen M.R., de Bruijn M.F., Liu P, & Göttgens B. (2014). Single-cell analyses of regulatory network perturbations using enhancer-targeting TALEs suggest novel roles for PU.1 during haematopoietic specification. Development (Cambridge, England), 141(20), 4018-4030.
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3

Immunofluorescence Staining of Cultured Cells

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Cells were rinsed 1× in prewarmed PBS and fixed by incubating in prewarmed 4% paraformaldehyde for 15 min at 37°C. Cells were then rinsed 3× for 5 min with PBS and permeabilized in 0.1% Triton X-100 for 15 min at RT. Cells were then rinsed with PBS 3× for 5 min and then blocked in 5% BSA for 1 h at 37°C. Following blocking, cells were briefly rinsed 1× with PBS and incubated with 1° antibodies for 1 h at 37°C (anti-HA rabbit, Invitrogen H6908; anti-myc rabbit; Abcam ab9106) or Alexa Fluor-488 conjugated streptavidin (Invitrogen S32354), both at 1:1000 dilution. Cells were then rinsed 4× for 5 min with PBS and incubated with 2° antibodies (Alexa Fluor 647 F(ab’)2-goat anti-rabbit IgG (H+L) A21246, 1:1000 and Alexa-Fluor 568 Phalloidin (Invitrogen A12380)), 1:200 for 30 min at RT. Following 2° antibody incubation, cells were rinsed 4× for 5 min in PBS and mounted on glass slides using ProLong Gold antifade reagent (Life Technologies, Invitrogen, P36930).
Gaeta I.M, & Tyska M.J. (2023). BioID2 screening identifies KIAA1671 as an EPS8 proximal factor that marks sites of microvillus growth. Molecular Biology of the Cell, 34(4), ar31.
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4

Western Blot Analysis of Cellular Proteins

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Proteins in cell lysates were separated by electrophoresis and transferred to a nitrocellulose Amersham Protran membrane (GE Healthcare). The membrane was blocked for 1 hr in 5% milk in PBS containing 0.1% Tween-20 at 23°C. Then, the membrane was incubated with primary antibody (Ab) diluted in blocking buffer at 4°C overnight. Antibodies used were from the following sources: DNA-PKcs (NeoMarkers, MS-423-P1, diluted 1:1000), Ku70 (AbCam, ab3114, diluted 1:1000), Ku80 (Santa Cruz, sc-1484, 1:500), FLAG (Sigma-Aldrich, F7425, diluted 1:5000), HA (Sigma-Aldrich, H6908, diluted 1:1000), P-IRF3 (Abcam, ab76493, diluted 1:1500), and α-tubulin (Merck Millipore, 05-829, diluted 1:10000). Membranes were probed and visualized with LI-COR Biosciences secondary Abs and the Li-Cor Odyssey infrared imaging system according to the manufacturer’s instructions. A quantification of protein bands was performed, where indicated, by using Odyssey software (LI-COR Biosciences).
Scutts S.R., Ember S.W., Ren H., Ye C., Lovejoy C.A., Mazzon M., Veyer D.L., Sumner R.P, & Smith G.L. (2018). DNA-PK Is Targeted by Multiple Vaccinia Virus Proteins to Inhibit DNA Sensing. Cell Reports, 25(7), 1953-1965.e4.
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5

Immunofluorescence and Western Blotting Protocol

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Antibodies used for immunofluorescence include anti-pH3 (Abcam, ab14955; 1:1,000), anti-Tbr1 (Abcam, ab31940; 1:300), anti-mCherry (Abcam, ab167453; 1:1,000), anti-NeuN (Millipore, MAB377; 1:300), anti-HA (Sigma-Aldrich, H6908; 1:2,000), anti-FLAG (Abcam, ab1162; 1:2,000), anti-LIC1 (Tan et al., 2011 (link); 1:300), and anti-LIC2 (Tynan et al., 2000a (link); 1:300). Donkey fluorophore–conjugated secondary antibodies (Jackson Labs, 1:500 dilution) were used.
Antibodies used for Western blotting include anti-p150Glued (BD, 610474; 1:1,000), anti–dynein IC clone 74.1 (University of Virginia, Charlottesville, VA; 1:1,000), anti-LICs (Tynan et al., 2000a (link); 1:500), anti-LIC1 (Tan et al., 2011 (link); 1:500), anti-LIC2 (Tynan et al., 2000a (link); 1:1,000), anti–dynein HC (Suzuki et al., 2007 (link); 1:1,000), and anti-GAPDH (Abcam, ab8245; 1:1,000). To develop in a LI-COR system, fluorescent secondary antibodies (1:10,000) were acquired from Invitrogen and Rockland.
Gonçalves J.C., Dantas T.J, & Vallee R.B. (2019). Distinct roles for dynein light intermediate chains in neurogenesis, migration, and terminal somal translocation. The Journal of Cell Biology, 218(3), 808-819.
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6

Extraction and Western Blot Analysis of Arabidopsis Proteins

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Total proteins from Arabidopsis plants were extracted by centrifugation and resuspension in buffer, before Western Blot24 (link), with the following antibodies: anti-HA (Sigma Aldrich; H6908; 1:10,000), anti-GUS (Abcam; ab50148; 1:5,000), anti-LUC (Agrisera; AS16 3691 A; 1:1,000), anti-Ein2 (Agrisera; AS12 1865; 1:10,000; also recognize SKL from M. truncatula), anti-Bak1 (Agrisera; AS12 1858; 1:2,000), anti-Bri1 (Agrisera; AS12 1859; 1:5,000), anti-DCL1 (Agrisera; AS19 4307; 1:1,000), anti-HSP101 (Agrisera; AS07 253; 1:1,000) anti-GFP (Sigma Aldrich; SAB4301138; 1:5,000). Anti-CPK3 and anti-GAPC antibodies were raised in rabbits by Covalab (France) using purified recombinant CPK3-GST and purified recombinant AtGAPC1-GST, respectively. Anti-CPK3 antibodies were purified from rabbit serum by affinity chromatography on 6His-CPK3 coupled to CH-sepharose (GE Healthcare), while crude serum was used for anti-GAPC western blots. Anti-CPK3 and anti-GAPC were used at a dilution of 1:5000 and 1:50,000, respectively. Protein levels were normalized to Ponceau staining.
Ormancey M., Guillotin B., Merret R., Camborde L., Duboé C., Fabre B., Pouzet C., Impens F., Van Haver D., Carpentier M.C., Clemente H.S., Aguilar M., Lauressergues D., Scharff L.B., Pichereaux C., Burlet-Schiltz O., Bousquet-Antonelli C., Gevaert K., Thuleau P., Plaza S, & Combier J.P. (2023). Complementary peptides represent a credible alternative to agrochemicals by activating translation of targeted proteins. Nature Communications, 14, 254.
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7

Protein Extraction and Western Blot Analysis

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Cells were lysed in 50 mM Tris–HCl (pH 7.4), 150 mM NaCl, 1 mM EDTA, 1% (v/v) Triton X‐100 and 1 × proteinase inhibitors for 30 min at 4°C, and equal protein amounts were separated on 4–20% Criterion TGX SDS–PAGE gels (Bio‐Rad). Primary antibodies were incubated overnight at 4°C, and secondary antibodies were incubated for 1 h at room temperature. Membranes were developed using Pierce ECL or SuperSignal West Pico Plus (Thermo Fisher). Antibodies used were as follows: rabbit anti‐HA (Sigma Aldrich H6908; RRID:AB_260070, 1:1,000), mouse anti‐β‐actin (Abcam ab6276; RRID:AB_2223210, 1:50,000), rabbit anti‐TOP3A (Proteintech 14525‐1‐AP; RRID:AB_2205881, 1:1,000), rabbit anti‐mouse HRP (Agilent P0260; RRID:AB_2636929, 1:3,000) and swine anti‐rabbit HRP (Agilent P0217; RRID:AB_2728719, 1:3,000).
Erdinc D., Rodríguez‐Luis A., Fassad M.R., Mackenzie S., Watson C.M., Valenzuela S., Xie X., Menger K.E., Sergeant K., Craig K., Hopton S., Falkous G., Poulton J., Garcia‐Moreno H., Giunti P., de Moura Aschoff C.A., Morales Saute J.A., Kirby A.J., Toro C., Wolfe L., Novacic D., Greenbaum L., Eliyahu A., Barel O., Anikster Y., McFarland R., Gorman G.S., Schaefer A.M., Gustafsson C.M., Taylor R.W., Falkenberg M, & Nicholls T.J. (2023). Pathological variants in TOP3A cause distinct disorders of mitochondrial and nuclear genome stability. EMBO Molecular Medicine, 15(5), e16775.
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8

Comprehensive Antibody Reference for Research

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The following antibodies were used in this study: anti-b-Actin (mouse monoclonal, Sigma-Aldrich, clone AC-15, A5441, 1:10,000), Anti-EGFR (rabbit monoclonal, Abcam, ab52894, 1:1000), Anti-Gephyrin (mouse monoclonal, Synaptic Systems, clone mAb7a, 147 021, 1:1000) Anti-GFP (chicken polyclonal, Abcam, ab13970, 1:600), Anti-GFP (rat monoclonal, ChromoTek, 3H9, 1:5000), Anti-GluA2 (rabbit polyclonal, Synaptic Systems, 182 103, 1:1000), Anti-GST (goat polyclonal, GE Healthcare, 27-4577-01, 1:10,000), Anti-HA (mouse monoclonal, Sigma-Aldrich, clone HA-7, H3663, 1:1000), Anti-HA (rabbit polyclonal, Sigma Aldrich, H6908, 1:100), Anti-LAMP1 (rabbit polyclonal, Abcam, ab24170, 1:1000), Anti-NLG1 (mouse monoclonal, Neuromab, N97A/31, 1:500), Anti-NLG2 (mouse monoclonal, Synaptic Systems, clone 5E6, 129 511, 1:2000 WB, 1:600 ICC), Anti-NLG3 (mouse monoclonal, Neuromab, N110/29, 1:500), Anti-GluN1 (rabbit monoclonal, Cell Signaling, D65B7 5704, 1:2000), Anti-SNX27 (mouse monoclonal, Abcam, clone 1C6, ab77799, 1:1000), Anti-SNX27 (rabbit polyclonal, a kind gift from Dr Martin Playford (NIH, USA), 1:2000), Anti-VPS35 (rabbit polyclonal, Abcam, ab97545, 1:2000).
Binda C.S., Nakamura Y., Henley J.M, & Wilkinson K.A. (2019). Sorting nexin 27 rescues neuroligin 2 from lysosomal degradation to control inhibitory synapse number. The Biochemical journal, 476(2).
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