17-677
ChIPAb+ Dimethyl-Histone H3 (Lys4) - ChIP Validated Antibody and Primer Set
clone CMA303, from mouse
Synonym(s):
H3K4me2, Histone H3 (di methyl K4)
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| Size/SKU | Availability | Price |
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25 assays | Check Cart for Availability | $648.00 |
About This Item
UNSPSC Code:
12352203
NACRES:
NA.52
eCl@ss:
32160702
Clone:
CMA303, monoclonal
Species reactivity:
human, vertebrates
Application:
ChIP, DB, WB, multiplexing
Citations:
15
biological source
mouse
Quality Segment
antibody form
purified immunoglobulin
clone
CMA303, monoclonal
species reactivity
human, vertebrates
manufacturer/tradename
ChIPAb+, Upstate®
technique(s)
ChIP: suitable (ChIP-seq), dot blot: suitable, multiplexing: suitable, western blot: suitable
isotype
IgG2bκ
NCBI accession no.
UniProt accession no.
shipped in
dry ice
General description
All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ Dimethyl-Histone H3 (Lys4) set includes the Anti-dimethyl-Histone H3 (Lys4) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 213 bp region within the coding region of the human GAPDH gene. The dimethyl-histone H3 (Lys4) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of dimethyl-histone H3 (Lys4)-associated chromatin.
The ChIPAb+ Dimethyl-Histone H3 (Lys4) set includes the Anti-dimethyl-Histone H3 (Lys4) antibody, a negative control antibody (purified Mouse IgG), and qPCR primers which amplify a 213 bp region within the coding region of the human GAPDH gene. The dimethyl-histone H3 (Lys4) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of dimethyl-histone H3 (Lys4)-associated chromatin.
Dimethyl-histone H3 at ~17 kDa
Immunogen
Epitope: a.a. 1-12
The dimethyl-histone H3 (Lys4) purified antibody is made against a synthetic peptide (dimethylated at Lys4) corresponding to amino acids 1-12 of histone H3.
Application
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either normal mouse IgG, or Anti-dimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of dimethyl-histone H3 (Lys4)-associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus GAPDH Coding primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each antibody with the indicated primers.
Please refer to the EZ-Magna G ChIP
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP
Western Blot Analysis:
Recombinant Histone H3 (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-dimethyl Histone H3 (Lys4) (2 μg/mL).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. #AP124P) and a chemiluminescence detection system.
Dot Blot Analysis:
Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Anti-dimethyl H3 (Lys4) at 2.0ug/mL (1:500) dilution. Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.
Beadlyte
0.15 μg/ml of purified anti-dimethyl-Histone H3 (Lys4),
clone CMA303 was incubated with a cocktail of microspheres
conjugated to histone H3 peptides with the following modifications:
1. unmodified H3 (K4)
2. monomethyl H3 (K4)
3. dimethyl H3 (K4)
4. trimethyl H3 (K4)
5. trimethy H3 (K27)
Unbound antibody was then removed by filtration.
Peptide antibody complexes were incubated with a
PE-conjugated anti-mouse secondary antibody.
Fluorescence was read on a Luminex
instrument. Median Fluorescence intensity (MFI) is plotted.
Sonicated chromatin prepared from HeLa cells (2 X 106 cell equivalents per IP) was subjected to chromatin immunoprecipitation using 2 µg of either normal mouse IgG, or Anti-dimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of dimethyl-histone H3 (Lys4)-associated DNA fragments was verified by qPCR using β-globin ChIP Primers versus GAPDH Coding primers (Please see figures). Data is presented as percent input of each IP sample relative to input chromatin for each antibody with the indicated primers.
Please refer to the EZ-Magna G ChIP
ChIP-seq Analysis:
Chromatin immunoprecipitation was performed using the Magna ChIP
Western Blot Analysis:
Recombinant Histone H3 (Lane 1) and HeLa acid extract (Lane 2) were resolved by electrophoresis, transferred to PVDF membrane and probed with Anti-dimethyl Histone H3 (Lys4) (2 μg/mL).
Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP (Cat. #AP124P) and a chemiluminescence detection system.
Dot Blot Analysis:
Absurance Histone H3 Antibody Specificity Array (Cat. No. 16-667) and Absurance Histone H2A, H2B, H4 Antibody Specificity Array (Cat. No. 16-665), which contain histone peptides with various modifications were probed with Anti-dimethyl H3 (Lys4) at 2.0ug/mL (1:500) dilution. Proteins were visualized using a Donkey anti-mouse IgG conjugated to HRP and a chemiluminescence detection system.
Beadlyte
0.15 μg/ml of purified anti-dimethyl-Histone H3 (Lys4),
clone CMA303 was incubated with a cocktail of microspheres
conjugated to histone H3 peptides with the following modifications:
1. unmodified H3 (K4)
2. monomethyl H3 (K4)
3. dimethyl H3 (K4)
4. trimethyl H3 (K4)
5. trimethy H3 (K27)
Unbound antibody was then removed by filtration.
Peptide antibody complexes were incubated with a
PE-conjugated anti-mouse secondary antibody.
Fluorescence was read on a Luminex
instrument. Median Fluorescence intensity (MFI) is plotted.
Research Category
Epigenetics & Nuclear Function
Epigenetics & Nuclear Function
Research Sub Category
Chromatin Biology
Chromatin Biology
This ChIPAb+ Dimethyl-Histone H3 (Lys4) -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.
Biochem/physiol Actions
Recognizes histone H3, Mr 17 kDa, dimethylated at lysine 4.
The peptide sequence is identical in a wide range of animal and plant species, so broad cross-reactivity is expected.
Packaging
25 assays per kit, ~2μg per chromatin immunoprecipitation
Physical form
Anti-dimethyl-Histone H3 (Lys4) (mouse monoclonal IgG1, clone CMA303). One vial containing 50 μg of protein G purified antibody in 50 μL PBS containing 0.05% sodium azide. Store at -20°C.
Normal Mouse IgG. Two vials containing 25 μg purified Mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers GAPDH Coding. One vial containing 75 μL of 5 μM of each primer specific for the coding region human GAPDH. Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
Normal Mouse IgG. Two vials containing 25 μg purified Mouse IgG in 25 μL storage buffer containing 0.1% sodium azide. Store at -20°C.
ChIP Primers GAPDH Coding. One vial containing 75 μL of 5 μM of each primer specific for the coding region human GAPDH. Store at -20°C.
FOR: GGC TCC CAC CTT TCT CAT CC
REV: GGC CAT CCA CAG TCT TCT GG
Format: Purified
Preparation Note
Stable for 1 year at -20°C from date of receipt.
Aliquot upon initial thaw, avoid freeze thaw cycles.
Aliquot upon initial thaw, avoid freeze thaw cycles.
Analysis Note
Chromatin Immunoprecipitation:
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG, or Anti-dimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of dimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding (Please see figures).
Please refer to the EZ-Magna G ChIP
Sonicated chromatin prepared from HeLa cells (1 X 106 cell equivalents per IP) were subjected to chromatin immunoprecipitation using 2 µg of either a normal mouse IgG, or Anti-dimethyl-Histone H3 (Lys4) antibody and the Magna ChIP G Kit (Cat. #17-611). Successful immunoprecipitation of dimethyl-histone H3 (Lys4) associated DNA fragments was verified by qPCR using ChIP Primers GAPDH Coding (Please see figures).
Please refer to the EZ-Magna G ChIP
Control
Includes negative control mouse IgG antibody and control primers specific for human β-globin promoter.
Includes negative control mouse IgG antibody and control primers specific for human β-globin promoter.
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
1 of 1
This Item | |||
|---|---|---|---|
| description clone CMA303, from mouse | description clone CMA304, from mouse, purified by using protein G | description clone CMA307, from mouse | description clone CMA306, from mouse, purified by using protein G |
| clone CMA303, monoclonal | clone CMA304, monoclonal | clone CMA307, monoclonal | clone CMA306, monoclonal |
| species reactivity human, vertebrates | species reactivity human, vertebrates | species reactivity human, vertebrates | species reactivity vertebrates, human |
| antibody form purified immunoglobulin | antibody form - | antibody form - | antibody form purified immunoglobulin |
| biological source mouse | biological source mouse | biological source mouse | biological source mouse |
| shipped in dry ice | shipped in dry ice | shipped in dry ice | shipped in dry ice |
| technique(s) ChIP: suitable (ChIP-seq), multiplexing: suitable, dot blot: suitable, western blot: suitable | technique(s) ChIP: suitable (ChIP-seq), immunocytochemistry: suitable, western blot: suitable | technique(s) ChIP: suitable, western blot: suitable | technique(s) ChIP: suitable (ChIP-seq), immunoprecipitation (IP): suitable, western blot: suitable |
Storage Class
10 - Combustible liquids
wgk
WGK 2
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