Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)
![Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)](https://www.abcam.com/ps/products/256/ab256361/Images/ab256361-369926-mousemonoclonals96todnarnahybriddriprloop.jpg "Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [S9.6] to DNA:RNA hybrid - BSA and Azide free
- Suitable for: IP, Dot blot
- Reacts with: Species independent
Overview
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Product name
Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free
See all DNA:RNA hybrid primary antibodies -
Description
Mouse monoclonal [S9.6] to DNA:RNA hybrid - BSA and Azide free -
Host species
Mouse -
Tested applications
Suitable for: IP, Dot blotmore details
Unsuitable for: ICC/IF or IHC-P -
Species reactivity
Reacts with: Species independent -
Immunogen
Chemical/ Small Molecule. This information is considered to be commercially sensitive.
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Positive control
- DRIP: R-Loop. Dot blot: R-Loop.
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General notes
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The pCALM3_2 vector used to generate R-loops was kindly provided by Prof. Frederic Chedin at UC Davis. The pCALM3_2 plasmid carries a 789bp fragment of the human CALM3 region that forms R-loops (PubMed ID: 31053798) cloned between T3 and T7 RNA polymerase promoter sequences. Transcription by T3 RNA polymerase leads to R-loop formation. Transcription by T7 RNA polymerase does not allow R-loop formation.
Our DNA-RNA immunoprecipitation (DRIP) protocol is linked here.
ab256361 is the carrier-free version of ab234957. 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
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.
Maxpar® is a trademark of Fluidigm Canada Inc.
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... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
S9.6 -
Isotype
IgG2a -
Light chain type
kappa
Images
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Immunoprecipitation - Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)
DNA-RNA hybrid Immunoprecipitation (DRIP) data using ab234957.
M1 : 100bp DNA ladder
M2 : High Range DNA Ladder
Lane 1: R-Loop ApaLI (RNase A treated)
Lane 2: R-Loop (RNase A treated)
Lane 3:R-Loop (RNase A+H treated)
Lane 4: R-Loop ApaLI (RNase A+H treated)
Lane 5: ab234957 with R-Loop ApaLI (RNase A treated)
Lane 6: ab234957 with R-Loop(RNase A treated)
Lane 7: ab234957 with R-Loop(RNase A+H treated)
Lane 8: ab234957 with R-Loop ApaLI (RNase A+H treated)
Lane 9-12: Flow through from IP
Capture antibody: 5 μg
Blocking buffer and concentration: 7.5 µg ssDNA.
Diluting buffer and concentration: PBS containing 0.1% Triton X-100.
A synthetic vector (pCALM3_2) was used to generate R-loops. ab234957 immunoprecipitates R-loops in the presence or absence of prior digestion by ApaLI, which does not affect R-loop structure. Prior treatment with RNase A (digests single stranded RNA) does not affect the IP signal whereas prior treatment with RNase H (digests RNA in DNA-RNA hybrids) eliminates the signal.
The pCALM3_2 vector used to generate R-loops was kindly provided by Prof. Frederic Chedin at UC Davis.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab234957).
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![Dot Blot - Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)](https://www.abcam.com/ps/products/256/ab256361/Images/ab256361-369924-mousemonoclonals96todnarnahybriddotblot.jpg)
Dot Blot - Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)Dot blot analysis with ab234957 at 1/1000 dilution.
Lane 1: Untranscribed plasmid.
Lane 2: R-Loop.
Lane 3: R-Loop (RNase A treated).
Lane 4: R-Loop (RNase A+H treated).
Lane 5: R-Loop (DNase I treated).
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution was used as secondary antibody.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
pCALM3_2 was cloned in pFC53 in the T7 orientation, but it forms R-loops transcribed in the reverse direction (that is with T3 promoter). DNase I has been shown to be less effective at digesting DNA in DNA-RNA hybrids compared to double stranded DNA (PubMed ID: 9020884). Prior treatment with DNase I reduced the signal by dot blot but did not eliminate it.
The pCALM3_2 vector used to generate R-loops was kindly provided by Prof. Frederic Chedin at UC Davis.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab234957).
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![Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)](https://www.abcam.com/ps/products/256/ab256361/Images/ab256361-3-benefits-of-recombinant-antibodies.png)
Anti-DNA:RNA hybrid antibody [S9.6] - BSA and Azide free (ab256361)
