Recombinant Anti-CRISPR-Cas9 antibody [EPR18991] - BSA and Azide free (ab232379)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18991] to CRISPR-Cas9 - BSA and Azide free
- Suitable for: ICC, Flow Cyt (Intra), WB, ICC/IF, IHC-P
Related conjugates and formulations
Overview
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Product name
Anti-CRISPR-Cas9 antibody [EPR18991] - BSA and Azide free
See all CRISPR-Cas9 primary antibodies -
Description
Rabbit monoclonal [EPR18991] to CRISPR-Cas9 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC, Flow Cyt (Intra), WB, ICC/IF, IHC-Pmore details -
Species reactivity
Predicted to work with: Streptococcus pyogenes -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB and Flow Cyt (intra): HEK-293 whole cell lysate transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag.IHC: 293T cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc). ICC/IF: 293T cells transfected with CRISPR-Cas9 with GFP-tag.
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General notes
ab232379 is the carrier-free version of ab189380.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. 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.
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
EPR18991 -
Isotype
IgG
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab232379 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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ICC |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 184 kDa (predicted molecular weight: 158 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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ICC
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 184 kDa (predicted molecular weight: 158 kDa). |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Target
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Relevance
[FUNCTION] CRISPR (clustered regularly interspaced short palindromic repeat) is an adaptive immune system that provides protection against mobile genetic elements (viruses, transposable elements and conjugative plasmids). CRISPR clusters contain spacers, sequences complementary to antecedent mobile elements, and target invading nucleic acids. CRISPR clusters are transcribed and processed into CRISPR RNA (crRNA) (Probable). In type II CRISPR systems correct processing of pre-crRNA requires a trans-encoded small RNA (tracrRNA), endogenous ribonuclease 3 (rnc) and this protein. The tracrRNA serves as a guide for ribonuclease 3-aided processing of pre-crRNA. Subsequently Cas9/crRNA/tracrRNA endonucleolytically cleaves linear or circular dsDNA target complementary to the spacer. The target strand not complementary to crRNA is first cut endonucleolytically, then trimmed by 3'-5' exonucleolytically. DNA-binding requires protein and both RNA species. Cas9 probably recognizes a short motif in the CRISPR repeat sequences (the PAM or protospacer adjacent motif) to help distinguish self versus nonself. -
Database links
- Entrez Gene: 901176 Streptococcus pyogenes
- SwissProt: Q99ZW2 Streptococcus pyogenes
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Alternative names
- Cas9 antibody
- CRISPR-associated endonuclease Cas9/Csn1 antibody
- CRISPR-Cas9/Csn1 antibody
see all
Images
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Intracellular Flow Cytometry analysis of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag) labelling CRISPR-Cas9 (red) with ab189380 at dilution of 1/70. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde. Isotype control antibody was (ab172730) Rabbit monoclonal IgG (black).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
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Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc) labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Positive staining on 293T cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc) 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
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Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with blank pcDNA3.1(+)-GFP-Myc vector labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Negative on 293T cells transfected with blank pcDNA3.1(+)-GFP-Myc vector.
Counter stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
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Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
No staining on Human stomach is observed.
Counter stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
No staining on mouse stomach is observed.
Counter stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
No staining on rat cerebrum is observed.
Counter stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (Human epithelial cell line from embryonic kidney) cells transfected with CRISPR-Cas9-GFP or GFP only, labeling CRISPR-Cas9 with ab189380 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 647) (ab150079) secondary antibody at 1/1000 dilution.
Confocal image showing positive staining on 293T cells transfected with CRISPR-Cas9 with GFP-tag.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab189380 at 1/500 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150079 (Alexa Fluor® 647 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab189380).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab232379 has not yet been referenced specifically in any publications.