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Phospho-Histone H3-S28 Rabbit pAb (AP0839)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat, Other (Wide Range Predicted)

Western blot analysis of various lysates using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at 1:1000 dilution. Both HeLa cells and NIH/3T3 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1s.

Western blot analysis of lysates from HeLa cells, using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at 1:900 dilution. HeLa cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from C6 cells, using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at 1:900 dilution. C6 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from NIH/3T3 cells, using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at 1:900 dilution. NIH/3T3 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes after serum-starvation overnight.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 0.5s.

ABclonal:Immunofluorescence - Phospho-Histone H3-S28 Rabbit pAb (AP0839)

Immunofluorescence analysis of H9C2 cells using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at dilution of 1:100. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of NIH/3T3 cells using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at dilution of 1:100. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of U2OS cells using Phospho-Histone H3-S28 Rabbit pAb (AP0839) at dilution of 1:100. Blue: DAPI for nuclear staining.

Overview

Product name	Phospho-Histone H3-S28 Rabbit pAb
Catalog No.	AP0839
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG

Background

Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. This structure consists of approximately 146 bp of DNA wrapped around a nucleosome, an octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures. This gene is intronless and encodes a replication-dependent histone that is a member of the histone H3 family. Transcripts from this gene lack polyA tails; instead, they contain a palindromic termination element. This gene is found in the large histone gene cluster on chromosome 6p22-p21.3.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S28 of human Histone H3 (NP_003520.1).
Sequence	ARKSA
Gene ID
Swiss Prot
Synonyms	H3/A; H3C2; H3C3; H3C4; H3C6; H3C7; H3C8; H3FA; H3C10; H3C11; H3C12; HIST1H3A; Phospho-Histone H3-S28
Calculated MW	16kDa
Observed MW	17kDa

Applications

Reactivity	Human, Mouse, Rat, Other (Wide Range Predicted)
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:2000 IF/ICC 1:50 - 1:200 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 50% glycerol, pH7.3.
Key application	Western blotting Immunofluorescence
Positive samples	HeLa treated by Calyculin A, NIH/3T3 treated by Calyculin A, C6 treated by Calyculin A
Cellular location	Chromosome, Nucleus.