Anti-Histone H3 (acetyl K122) antibody

• ICC/IF

AbReview27812****

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K122) antibody (AB33309)

Immunocytochemical analysis of Arabidopsis thaliana root cells, labeling Histone H3 (acetyl K122) with ab33309. Cells were paraformaldehyde fixed and blocked in 3% BSA for 1 hour at 4°C. Incubation with ab33309 (diluted 1/1000) was for 18 hours at 4°C.

This image is courtesy of an anonymous Abreview.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (acetyl K122) antibody (AB33309)

ICC/IF image of ab33309 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab33309, 1μg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).

• WB

Project2250****

Western blot - Anti-Histone H3 (acetyl K122) antibody (AB33309)

ab33309 shows very slight cross reactivity with Histone H4 in western blot.

All lanes:

Western blot - Anti-Histone H3 (acetyl K122) antibody (ab33309) at 1 µg/mL

Lane 1:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg

Lane 2:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg with Human Histone H3 (acetyl K122) peptide (<a href='/en-us/products/unavailable/human-histone-h3-acetyl-k122-peptide-ab34466'>ab34466</a>)

Lane 3:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg with Histone H3 peptide - unmodified (ab34467)

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 11.2 kDa,17 kDa

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• WB

Lab

Western blot - Anti-Histone H3 (acetyl K122) antibody (AB33309)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab33309 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

All lanes:

Western blot - Anti-Histone H3 (acetyl K122) antibody (ab33309) at 1 µg/mL

Lane 1:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg

Lane 2:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg with Human Histone H3 (acetyl K122) peptide (<a href='/en-us/products/unavailable/human-histone-h3-acetyl-k122-peptide-ab34466'>ab34466</a>)

Lane 3:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg with Histone H3 peptide - unmodified (ab34467)

Lane 4:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg with Human Histone H3 (acetyl K56) peptide

Lane 5:

HeLa Histone Preparation Nuclear Lysate - Butyrated at 10 µg with Histone H3 peptide - unmodified K56 (ab73002)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/10000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

true

Exposure time: 2min

• WB

CiteAb

Western blot - Anti-Histone H3 (acetyl K122) antibody (AB33309)

Histone H3 (acetyl K122) western blot using anti-Histone H3 (acetyl K122) antibody ab33309. Publication image and figure legend from Shi, G. X., Zheng, X. F., et al., 2017, Int J Mol Sci, PubMed 28272338.

ab33309 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab33309 please see the product overview.

Lgr4 was essential for Rspo1 to promote osteogenic differentiation and Wnt/β-catenin signaling activity in BMSCs. (A) qRT-PCR analysis of Lgr4 in distal femurs of Con and TS mice (n = 6) and (B) in BMSCs treated with 5% CMS; (C) Western blot and qRT- PCR analysis of Lgr4 after lentivirus sh-Lgr4 infection; (D) qRT-PCR analysis of osteogenic differentiation markers (Ocn, Col-1a1, ALP) and ALP activities assay of lentivirus infected BMSCs after three days of osteogenic induction with or without Rspo1 (100 ng/mL); (E) representative images of ALP staining (Day 10) and Alizarin red staining (Day 18) of each group cells; (F) Western blot analysis of the intranuclear level of β-catenin and (G) Topflash dual-luciferase activity assay of BMSCs after 24 h of osteogenic differentiation; (H) qRT-PCR analysis of the Wnt target genes Axin2 and (I) Tcf1. All data were confirmed by three repeated tests and are presented as the mean ± SD, * p < 0.05, ** p < 0.01. All p-values are based on Student's t-test. NC : negative control; sh-Lgr4 : short hairpin RNA-Lgr4

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