Antibodies, Proteins, Kits and Reagents for Life Science

328 339 ₸ Product size

100 µl 328 339 ₸

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Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17899] to Histone H3.3 - ChIP Grade
Suitable for: ChIP-sequencing, ICC/IF, ChIP, IHC-P, WB, Dot blot
Reacts with: Mouse, Rat, Human

Product name

Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade
See all Histone H3.3 primary antibodies
Description

Rabbit monoclonal [EPR17899] to Histone H3.3 - ChIP Grade
Host species

Rabbit
Specificity

The immunogen sequence used for this antibody is 100% identical to the protein sequence shown for H3.X and H3.Y in Szenker et al., 2011 (PubMed ID 21263457). These H3.X and H3.Y protein sequences have not been reviewed by UniProt. The immunogen sequense is also 100% homologous to Histone H3.6. For further information on this antibody, please contact our technical support team.

Tested Applications & Species

Application	Species
ChIP	Human
ChIP-seq	Human
Dot	Mouse
ICC/IF	Human
IHC-P	Mouse Rat Human
WB	Mouse Human

See all applications and species data

Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HeLa and NIH/3T3 whole cell lysates. IHC-P: Human colon, Mouse stomach and Rat colon tissues. ICC/IF: HeLa cells. ChIP: Chromatin prepared from HeLa cells. ChIP-seq: Chromatin prepared from HeLa cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17899
Isotype

IgG
Research areas

ChIP-sequencing - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Chromatin was prepared from HeLa cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 30 µg of chromatin and 4 µg of Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840). ChIP DNA was sequenced on the Illumina NextSeq 500 to a depth of 30 million reads. ChIP-Seq validation performed by Active Motif, Carlsbad, CA.

Additional screenshots of mapped reads can be downloaded here.
ChIP - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Chromatin was prepared from HeLa (Human epithelial cells from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25µg of chromatin, 2µg of ab176840 (blue), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Western blot - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

All lanes : Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Dot Blot - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Dot blot analysis of Histone H3.3 peptide (Lane 1), and Histone H3.1 peptide (Lane 2), labeled using ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated secondary antibody at 1/1000 dilution.
Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

All lanes : Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840) at 1/1000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with Histone H3.3 peptide
Lane 3 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with Histone H3.1 peptide
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lane 5 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with Histone H3.3 peptide
Lane 6 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate with Histone H3.1 peptide

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 15 kDa
Observed band size: 15 kDa

Exposure time: 3 minutes

Blocking experiment by pre-incubating Histone H3.3 peptide with the antibody in lanes 2 and 5 and Histone H3.1 peptide in lanes 3 and 6 to show the antibody specificity against Histone H3.3.
Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab176840 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Human colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on mouse stomach tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H3.3 with ab176840 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nucleus staining on Rat colon tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Histone H3.3 antibody [EPR17899] - ChIP Grade (ab176840)

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