Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17589] to Histone H2A.X+Histone H2A (acetyl K5) - BSA and Azide free
- Suitable for: PepArr, ChIP, WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free
See all Histone H2A.X+Histone H2A primary antibodies -
Description
Rabbit monoclonal [EPR17589] to Histone H2A.X+Histone H2A (acetyl K5) - BSA and Azide free -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ChIP HumanIHC-P MousePepArr HumanWB Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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General notes
Ab250006 is the carrier-free version of ab177863. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab250006 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17589 -
Isotype
IgG -
Research areas
Images
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ChIP - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
This data was developed using ab177863, the same antibody clone in a different buffer formulation.
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 ab177863(red), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach). Primers and probes are located in the first kb of the transcribed region. -
Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)All lanes : Anti-Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) antibody [EPR17589] - ChIP Grade (ab177863) at 1/4000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) treated with 500 ng/ml Trichostatin A for 4 hours. Whole cell lysates
Lane 2 : Untreated HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) treated with 500 ng/ml Trichostatin A for 4 hours. Whole cell lysates
Lane 4 : Untreated NIH/3T3 (Human epithelial cell line from cervix adenocarcinoma) whole cell lysates
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: 14,15 kDa
Observed band size: 14,15 kDa why is the actual band size different from the predicted?This data was developed using ab177863, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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Immunocytochemistry - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)
This data was developed using ab177863, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cell line labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cell line. Acetylation level increased after treatment with Trichostatin A (500 ng/ml) for 4 hours. The nuclear counter stain is DAPI (blue).Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab177863 at 1/400 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/500 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution. -
Peptide Array - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)This data was developed using ab177863, the same antibody clone in a different buffer formulation.ab177863 was tested in Peptide Array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here. -
Western blot - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)All lanes : Anti-Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) antibody [EPR17589] - ChIP Grade (ab177863) at 1/4000 dilution
All lanes : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell line, treated with 500 ng/ml Trichostatin A for 4 hours. 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
Developed using the ECL technique.
Predicted band size: 14,15 kDa
Observed band size: 14,15 kDa why is the actual band size different from the predicted?This data was developed using ab177863, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)This data was developed using ab177863, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on human colon tissue is observed. Counter stained with Hematoxylin. Negative 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)This data was developed using ab177863, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on mouse colon tissue is observed. Counter stained with Hematoxylin. Negative 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H2A.X+Histone H2A (acetyl K5) antibody [EPR17589] - BSA and Azide free (ab250006)This data was developed using ab177863, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling Histone H2A.X (acetyl K5) + Histone H2A (acetyl K5) with ab177863 at 1/400 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on rat colon tissue is observed. Counter stained with Hematoxylin. Negative 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.
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