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Phospho-Histone H3-S28 Rabbit mAb (AP1431)

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 lysates from HeLa cells, using Phospho-Histone H3-S28 Rabbit mAb (AP1431) at 1:1000 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: 30s.

Western blot analysis of lysates from NIH/3T3 cells, using Phospho-Histone H3-S28 Rabbit mAb (AP1431) at 1:1000 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: 30s.

Western blot analysis of lysates from C6 cells, using Phospho-Histone H3-S28 Rabbit mAb (AP1431) at 1:1000 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: 30s.

ABclonal:Immunoprecipitation - Phospho-Histone H3-S28 Rabbit mAb (AP1431)

Immunoprecipitation analysis of 600 μg extracts of HeLa cells using 5 μg Phospho-Histone H3-S28 Rabbit mAb (AP1431). Western blot was performed from the immunoprecipitate using Phospho-Histone H3-S28 Rabbit mAb (AP1431) at a dilition of 1:2000.

Overview

Product name	Phospho-Histone H3-S28 Rabbit mAb
Catalog No.	AP1431
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC60274

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.
Sequence	RKSAP
Gene ID
Swiss Prot
Synonyms	H3/A; H3C2; H3C3; H3C4; H3C6; H3C7; H3C8; H3FA; H3C10; H3C11; H3C12; HIST1H3A; Phospho-Histone H3-S28
Calculated MW	15kDa
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 - 1:1000 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunoprecipitation
Positive samples	HeLa treated by Calyculin A, NIH/3T3 treated by Calyculin A, C6 treated by Calyculin A
Cellular location	Chromosome, Nucleus
Customer validation	WB(Homo sapiens)