Antibodies, Proteins, Kits and Reagents for Life Science

307 566 ₸ Product size

100 µl 307 566 ₸ 10 µl 103 862 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR23614-91] to Histone H3 (mutated K36M)
Suitable for: IP, WB, IHC-P, ICC/IF, Flow Cyt, Indirect ELISA
Reacts with: Human

Product name

Anti-Histone H3 (mutated K36M) antibody [EPR23614-91]
See all Histone H3 primary antibodies
Description

Rabbit monoclonal [EPR23614-91] to Histone H3 (mutated K36M)
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
IP	Human Recombinant fragment
WB	Recombinant fragment

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag, whole cell lysate IHC-P: Human chondroblastoma tissue. ICC/IF: HEK-293 cells Flow Cyt: 293T transfected with myc tagged Histone H3 construct cell. IP: HEK-293 transfected with Histone H3.3 K36M cell.
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

pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23614-91
Isotype

IgG
Research areas

Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

All lanes : Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384) at 1/1000 dilution

Lane 1 : HEK-293 transfected with Histone H3.3 (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 15 kDa
Observed band size: 20 kDa
why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST

Exposure time: 26 seconds
Immunoprecipitation - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Histone H3.3 (mutated K36 M) was immunoprecipitated from 0.35 mg HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag^®, whole cell lysate with ab256384 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab256384 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1: HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate 10 ug

Lane 2: ab256384 IP in HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag®, whole cell lysate

Lane 3:Rabbit monoclonal IgG (ab172730) instead of ab256384 in HEK-293 transfected with Histone H3.3 K36M (mutate) expression vector containing a myc-His-tag^®, whole cell lysate

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 10 seconds
Flow Cytometry - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HEK-293T transfected with myc tagged Histone H3 construct (Left) or myc tagged Histone H3 K36M construct (Right) cells labelling Histone H3 (mutated K36 M) with ab256384 at 1/500 compared with a isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 was used as the secondary antibody.
Indirect ELISA - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Indirect ELISA using ab256384 at varying antibody concentrations and antigen concentration at 100 ng/mL. An Alkaline Phosphatase-conjugated Goat Anti-Rabbit IgG (H+L) (1/2500) was used as the secondary antibody.

Binding was seen for Histone H3 (mutated K36M) peptide. Binding to the following peptides was not seen:

Histone H3 WT,
Histone H3 (mono methyl K36),
Histone H3 (di methyl K36),
Histone H3 (tri methyl K36).

This indicates the specificity of ab256384 for mutated K36M of Histone H3.
Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293 cells labelling Histone H3 (mutated K36 M) with ab256384 at 1/50 dilution, followed by a secondary antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HEK-293 cell line transfected with myc-tagged Histone H3 K36M expression vector. 2233S Myc-Tag (9B11) Mouse mAb (Alexa Fluor® 647 Conjugate) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Immunohistochemical analysis of paraffin-embedded human giant cell tumor of bone tissue labeling Histone H3 (mutated K36 M) with ab256384 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Negative control: No staining in human giant cell tumor of bone (PMID: 29757500). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
Western blot - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

All lanes : Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384) at 1/1000 dilution

Lane 1 : HEK-293 transfected with Histone H3.1 (WT) expression vector containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293 transfected with Histone H3.1 K36M (mutant)
expression vector containing a myc-His-tag®, whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 15 kDa
Observed band size: 19 kDa why is the actual band size different from the predicted?

Exposure time: 62 seconds

Blocking/Diluting buffer and concentration: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Immunohistochemical analysis of paraffin-embedded human chondroblastoma tissue labeling Histone H3 (mutated K36 M) with ab256384 at 1/4000 dilution followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear staining in human chondroblastoma (PMID: 29757500). Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0)
Anti-Histone H3 (mutated K36M) antibody [EPR23614-91] (ab256384)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com