Histone H2B Recombinant Rabbit Monoclonal Antibody [SD20-63] - BSA and Azide free
Catalog# HA750287
Histone H2B Recombinant Rabbit Monoclonal Antibody [SD20-63] - BSA and Azide free
-
WB
-
IF-Tissue
-
IHC-P
-
IP
-
ChIP
-
Human
-
Mouse
-
Rat
-
unconjugated
Overview
Product Name
Histone H2B Recombinant Rabbit Monoclonal Antibody [SD20-63] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human Histone H2B aa 110 to the C-terminus.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Tissue, IHC-P, IP, ChIP
Target Molecular Weight
Predicted band size: 14 kDa
Positive Control
Hela cell lysate, PC-12 cell lysate, human tonsil tissue, human liver tissue, mouse liver tissue, mouse testis tissue, mouse colon tissue, human breast carcinoma tissue
Conjugation
unconjugated
Clone Number
SD20-63
Product Features
Form
Liquid
Concentration
1mg/ml
Storage Instructions
Store at 2-8℃. Avoid freeze.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
Target
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.
Background References
1. Hoedt E et al. SILAC-based proteomic profiling of the human MDA-MB-231 metastatic breast cancer cell line in response to the two antitumoral lactoferrin isoforms: the secreted lactoferrin and the intracellular delta-lactoferrin. PLoS One 9:e104563 (2014).
2. Schmitz M et al. Impact of the cellular prion protein on amyloid- and 3PO-tau processing. J Alzheimers Dis 38:551-65 (2014).
Sequence Similarity
Belongs to the histone H2B family.
Post-translational Modification
Monoubiquitination at Lys-35 (H2BK34Ub) by the MSL1/MSL2 dimer is required for histone H3 'Lys-4' (H3K4me) and 'Lys-79' (H3K79me) methylation and transcription activation at specific gene loci, such as HOXA9 and MEIS1 loci. Similarly, monoubiquitination at Lys-121 (H2BK120Ub) by the RNF20/40 complex gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation. It also functions cooperatively with the FACT dimer to stimulate elongation by RNA polymerase II. H2BK120Ub also acts as a regulator of mRNA splicing: deubiquitination by USP49 is required for efficient cotranscriptional splicing of a large set of exons.; Phosphorylation at Ser-37 (H2BS36ph) by AMPK in response to stress promotes transcription (By similarity). Phosphorylated on Ser-15 (H2BS14ph) by STK4/MST1 during apoptosis; which facilitates apoptotic chromatin condensation. Also phosphorylated on Ser-15 in response to DNA double strand breaks (DSBs), and in correlation with somatic hypermutation and immunoglobulin class-switch recombination.; GlcNAcylation at Ser-113 promotes monoubiquitination of Lys-121. It fluctuates in response to extracellular glucose, and associates with transcribed genes (By similarity).; Crotonylation (Kcr) is specifically present in male germ cells and marks testis-specific genes in post-meiotic cells, including X-linked genes that escape sex chromosome inactivation in haploid cells. Crotonylation marks active promoters and enhancers and confers resistance to transcriptional repressors. It is also associated with post-meiotically activated genes on autosomes.
Subcellular Location
Nucleus, Chromosome.
UNIPROT
Synonyms
H2B GL105 antibody
H2B histone family member O antibody
H2B histone family member S antibody
H2B.1 antibody
H2B.1 B antibody
H2B.b antibody
H2B.c antibody
H2B.d antibody
H2B.e antibody
H2B.f antibody
ExpandImages
-
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
ICC staining of Histone H2B in A431 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 (ET1612-25, 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). -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Histone H2B antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1612-25, 1/50) 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. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Histone H2B antibody (ET1612-25) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-25) at 1/5,000 dilution for 1 hour 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. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-Histone H2B antibody (ET1612-25) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-25) at 1/5,000 dilution for 1 hour 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. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Histone H2B antibody (ET1612-25) at 1/2,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-25) at 1/2,000 dilution for 1 hour 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. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-Histone H2B antibody (ET1612-25) at 1/5,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-25) at 1/5,000 dilution for 1 hour 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. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Histone H2B antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1612-25, 1/50) 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. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of Histone H2B was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1612-25, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells with Histone H2B (ET1612-25) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one. -
This data was developed using ET1612-25, the same antibody clone in a different buffer formulation.
Western blot analysis of Histone H2B on different lysates with Rabbit anti-Histone H2B antibody (ET1612-25) at 1/1,000 dilution.
Lane 1: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
Lane 2: COS-1 (African green monkey kidney fibroblast) cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 77 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ET1612-25, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 14 kDa
Observed band size: 14 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"