PRODUCT CODE: ET1602-11

Phospho-Histone (H1.3(T17)+H1.4(T17)) Recombinant Rabbit Monoclonal Antibody [SR38-03] (ET1602-11)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Phospho-Histone H1.3(T17)+Histone H1.4(T17) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Untreated CRC whole cell lysates<br />
Lane 2: CRC cells treated with 1.5ug/ml Colcemid for 12 hours whole cell lysates
  • Western blot analysis of Phospho-Histone H1.3(T17)+Histone H1.4(T17) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: Untreated CRC whole cell lysates<br />
Lane 2: CRC cells treated with 1.5ug/ml Colcemid for 12 hours whole cell lysates
  • ICC staining of Phospho-Histone H1.3(T17)+Histone H1.4(T17) in NIH/3T3 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-11, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Phospho-Histone H1.3(T17)+Histone H1.4(T17) in CRC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1602-11, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded mouse colon cancer tissue using anti-Phospho-Histone H1.3(T17)+Histone H1.4(T17) antibody. Counter stained with hematoxylin.
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Phospho-Histone H1.3(T17)+Histone H1.4(T17) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-11, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-Phospho-Histone H1.3(T17)+Histone H1.4(T17) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-11, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-Histone H1.3(T17)+Histone H1.4(T17) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1602-11, 1/200)  for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Western blot analysis of Phospho-Histone H1.3(T17)+Histone H1.4(T17) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1602-11, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:40,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: Untreated CRC whole cell lysates
Lane 2: CRC cells treated with 1.5ug/ml Colcemid for 12 hours whole cell lysates

Applications

  • WB

  • ICC

  • IF

  • IHC-P

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Phospho-Histone (H1.3(T17)+H1.4(T17)) Recombinant Rabbit Monoclonal Antibody [SR38-03] (ET1602-11)

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding thr17 of human h1.4.

Host

Rabbit

Modification

Phospho

Modification Site

H1.3(T17)+H1.4(T17)

Positive Control

NIH/3T3, CRC, human colon cancer tissue, mouse colon cancer tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SR38-03

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A purified.

MOLECULAR WEIGHT

30 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:1,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:200

TARGET

UNIPROT #

PROTEIN NAME

Phospho-Histone (H1.3(T17)+H1.4(T17))

SYNONYMS

Histone H1.3; Histone H1c; Histone H1s-2; HIST1H1DH1F3; Histone H1.4; Histone H1b; Histone H1s-4; HIST1H1EH1F4

SEQUENCE SIMILARITIES

Belongs to the histone H1/H5 family.

POST-TRANSLATIONAL MODIFICATION

H1 histones are progressively phosphorylated during the cell cycle, becoming maximally phosphorylated during late G2 phase and M phase, and being dephosphorylated sharply thereafter.; Acetylated at Lys-26. Deacetylated at Lys-26 by SIRT1.; Citrullination at Arg-54 (H1R54ci) by PADI4 takes place within the DNA-binding site of H1 and results in its displacement from chromatin and global chromatin decondensation, thereby promoting pluripotency and stem cell maintenance.; ADP-ribosylated on Ser-150 in response to DNA damage.

SUBCELLULAR LOCATION

Nucleus, Chromosome

FUNCTION

Eukaryotic histones are basic and water soluble nuclear proteins that form hetero-octameric nucleosome particles by wrapping 146 base pairs of DNA in a left-handed super-helical turn sequentially to form chromosomal fiber. Two molecules of each of the four core histones (H2A, H2B, H3, and H4) form the octamer; formed of two H2A-H2B dimers and two H3-H4 dimers, forming two nearly symmetrical halves by tertiary structure. Over 80% of nucleosomes contain the linker Histone H1, derived from an intronless gene, that interacts with linker DNA between nucleosomes and mediates compaction into higher order chromatin. Histones are subject to posttranslational modification by enzymes primarily on their N-terminal tails, but also in their globular domains. Such modifications include methylation, citrullination, acetylation, phosphorylation, sumoylation, ubiquitination and ADP-ribosylation.