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Ab31830

Manufactured by Abcam
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Ab31830 is a primary antibody that recognizes the Glial fibrillary acidic protein (GFAP) in human, mouse, and rat samples. GFAP is an intermediate filament protein that is widely used as a marker for astrocytes in the central nervous system.

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Lab products found in correlation

Ab17677, by Abcam (1 mentions) Ab18208, by Abcam (1 mentions) Ab24834, by Abcam (1 mentions) Anti h3, by Active Motif (1 mentions) Anti h2a, by Merck Group (1 mentions) Hmgn2, by Cell Signaling Technology (1 mentions) Atto 488 goat anti mouse igg, by Merck Group (1 mentions) Goat anti rabbit igg atto 488, by Merck Group (1 mentions) Normal goat serum (ngs), by Thermo Fisher Scientific (1 mentions) Alexa fluor 546 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 633 conjugated goat anti guinea pig igg, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 conjugated goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Ab9051, by Abcam (1 mentions) Sc 56198, by Santa Cruz Biotechnology (1 mentions) Ab1791, by Abcam (1 mentions) Acetyl histone h3, by Merck Group (1 mentions) Acetyl histone h4, by Merck Group (1 mentions)

5 protocols using ab31830

1

Antibody Immunostaining Protocol

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The following primary rabbit antibodies were purchased from Abcam: anti-H1.2, ab17677; H1.5, ab18208; HMGB2, ab124670; Ki67, ab15580; H3S10p, ab5176; CTCF, ab128873; SMC2, ab10412; RAD21, ab154769; H1x, ab31972. From Cell Signaling: anti-HMGN1, #5692; HMGN2, #9437. From Sigma-Aldrich: anti-H1.4, H7665. From Millipore: anti-H2A, #07-146. The following mouse antibodies were purchased from Abcam: anti-H3, ab24834; H4, ab31830; H2B, ab52584. From Active Motif: anti-H3, #61475. mAb PL2-6 was a gift from M. Monestier (Temple Univ.), see previous use [26,27]. Secondary antibodies included Invitrogen Alexa 568 goat anti-rabbit IgG, Sigma-Aldrich Atto 488 goat anti-rabbit IgG and Atto 488 goat anti-mouse IgG.
Olins A.L., Gould T.J., Boyd L., Sarg B, & Olins D.E. (2020). Hyperosmotic stress: in situ chromatin phase separation. Nucleus, 11(1), 1-18.
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2

Immunostaining of Histone H4 and Chromatin

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Lam-KD, LBR-KD or control S2 cells were seeded on coverslips for 30 min. After rinsing with PBS, cells were fixed in 100% methanol for 5 min at room temperature (for further examination of chromatin distribution based on the immunostaining of histone H4) or in 4% formaldehyde in PBS for 25 min at room temperature (for further estimation of chromatin volume based on DAPI staining), rinsed with PBS three times, blocked with PBTX (PBS with 0.1% Tween-20 and 0.3% Triton X-100) containing 3% normal goat serum (Invitrogen) for 1 h at room temperature. The remaining immunostaining procedure was performed as previously described50 (link). As primary antibodies we used murine monoclonal anti-histone H4 (1:200; ab31830, Abcam), guinea pig polyclonal anti-LBR26 (link) (1:1000), rabbit polyclonal anti-lamin Dm051 (link) (1:500). As the secondary antibodies we used Alexa Fluor 546-conjugated goat anti-rabbit IgG (Invitrogen) or Alexa Fluor 488-conjugated goat anti-mouse IgG (Invitrogen), or Alexa Fluor 633-conjugated goat anti-guinea pig IgG (Invitrogen).
Ulianov S.V., Doronin S.A., Khrameeva E.E., Kos P.I., Luzhin A.V., Starikov S.S., Galitsyna A.A., Nenasheva V.V., Ilyin A.A., Flyamer I.M., Mikhaleva E.A., Logacheva M.D., Gelfand M.S., Chertovich A.V., Gavrilov A.A., Razin S.V, & Shevelyov Y.Y. (2019). Nuclear lamina integrity is required for proper spatial organization of chromatin in Drosophila. Nature Communications, 10, 1176.
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3

Immunoblotting Antibodies for Protein Analysis

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The following antibodies were used for immunoblotting assays: anti-PR-SET7 (Rabbit, 1:1000, Novus Biologicals, 44710002), anti-H4K20me1 (Rabbit, 1:1000, Abcam, ab9051), anti-H4 (Mouse, 1:1000, Abcam, ab31830), anti-pADPr (Mouse monoclonal, 1:500, 10 H - sc-56198, Santa Cruz), anti-H3 (Rabbit polyclonal, 1/1000, FL-136 sc-10809 Santa Cruz).
Bamgbose G., Bordet G., Lodhi N, & Tulin A. (2024). Mono-methylated histones control PARP-1 in chromatin and transcription. eLife, 13, RP91482.
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4

Histone Antibody Labeling Protocol

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Rabbit anti-histone H3 polyclonal antibody (ab1791), mouse anti-histone H4 monoclonal antibody (ab31830), goat polyclonal secondary antibody to rabbit IgG-Fc (DyLight 488) (ab98462), goat anti-mouse IgG H&L (Cy3) (ab97035), goat anti-rabbit IgG H&L (10 nm Gold) (ab39601), and goat anti-mouse IgG H&L (20 nm Gold) (ab27242) were purchased from Abcam (Cambridge, MA, USA). Horseradish peroxidase (HRP)-conjugated goat anti-rabbit, and HRP-conjugated goat anti-mouse antibodies were purchased from Beijing Solarbio Science&Technology Co., Ltd. (Beijing, P. R. China).
Wu J.L., Kang X.J., Guo M.S., Mu S.M, & Zhang Z.H. (2015). Cloning and Functional Analysis of Histones H3 and H4 in Nuclear Shaping during Spermatogenesis of the Chinese Mitten Crab, Eriocheir sinensis. PLoS ONE, 10(5), e0126623.
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5

Histone Acetylation Analysis

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Cells (107 cells per milliliter) were lysed in Triton Extraction Buffer (TEB) containing 0.5% Triton X-100 (vol/vol), 2 mM phenylmethylsulfonyl fluoride, and 0.02% (wt/vol) NaN3 in phosphate buffered saline (PBS) for 10 minutes on ice with gentle stirring. After centrifugation at 6,500g for 10 minutes at 4°C, the supernatant was discarded and the pellet was washed in half the volume of TEB and centrifuged as before. The pellet was resuspended in 0.2 N HCl at a cell density of 4 × 107 cells per milliliter and acid extraction was carried out overnight at 4°C. Then, the samples were centrifuged at 6,500g for 10 minutes at 4°C, the supernatant removed, and protein content determined using Bradford assay. Equal amounts of histones (10 µg) were separated on a 15% SDS-polyacrylamide gel and then transferred to a polyvinylidene difluoride membrane. Membranes were incubated with primary antibodies to acetyl-histone H3 (#06-599: Millipore, Vimodrone [MI], Italy, http://www.millipore.com.) and acetyl-histone H4 (#06-598: Millipore, Vimodrone, Mi, Italy). Total histone H4 (ab31830: Abcam, Cambridge U.K., http://www.abcam.com) antibody was used for assessing loading. Proteins were detected according to manufacturer's protocols and visualized using chemiluminescence (GE Healthcare U.K. Limited, U.K., http://www.gehealthcare.com).
Paino F., la Noce M., Tirino V., Naddeo P., Desiderio V., Pirozzi G., De Rosa A., Laino L., Altucci L, & Papaccio G. (2014). Histone Deacetylase Inhibition with Valproic Acid Downregulates Osteocalcin Gene Expression in Human Dental Pulp Stem Cells and Osteoblasts: Evidence for HDAC2 Involvement. Stem Cells (Dayton, Ohio), 32(1), 279-289.
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