Recombinant Anti-CRISPR-Cas9 antibody [KANI345B] (ab271293)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rat monoclonal [KANI345B] to CRISPR-Cas9
- Suitable for: WB, IHC-P, ICC
Related conjugates and formulations
Overview
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Product name
Anti-CRISPR-Cas9 antibody [KANI345B]
See all CRISPR-Cas9 primary antibodies -
Description
Rat monoclonal [KANI345B] to CRISPR-Cas9 -
Host species
Rat -
Tested applications
Suitable for: WB, IHC-P, ICCmore details -
Species reactivity
Predicted to work with: Streptococcus pyogenes -
Immunogen
Recombinant full length protein. This information is considered to be commercially sensitive.
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Positive control
- WB: HEK-293 transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag, whole cell lysate. IHC-P: HEK-293T transfected with a GFP-Myc-tagged CRISPR-associated endonuclease Cas9/Csn1 construct. ICC: HEK-293T cells transfected with GFP-tagged Cas9 expression vector.
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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.
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
Properties
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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
Preservative: 0.01% Sodium azide
Constituents: 59.04% PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine) -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
KANI345B -
Isotype
IgG2a
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry reagents
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab271293 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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WB |
1/5000.
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IHC-P | (1) |
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC |
1/50.
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Notes |
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WB
1/5000. |
IHC-P
1/200. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC
1/50. |
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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All lanes : Anti-CRISPR-Cas9 antibody [KANI345B] (ab271293) at 1/5000 dilution
Lane 1 : HEK-293 (human embryonic kidney epithelial cell) transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag, whole cell lysate
Lane 2 : HEK-293T transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 3 : HEK-293 transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag, whole cell lysate
Lane 4 : HEK-293 transfected with CRISPR-Cas9 (Q03JI6, Streptococcus thermophilus) with Myc-His tag, whole cell lysate
Lane 5 : HEK-293 transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag, whole cell lysate
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded (Panel A) HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a GFP-Myc-tagged CRISPR-associated endonuclease Cas9/Csn1 construct and (Panel B) HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with empty plasmid, labeling CRISPR-Cas9 with ab271293 at 1/200 (4.58 ug/ml) dilution followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Counterstained with Hematoxylin.
Positive staining on (A) HEK-293T transfected with a GFP-Myc-tagged CRISPR-associated endonuclease Cas9/Csn1 construct, no staining on (B) HEK-293T transfected with empty plasmid.Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T+OE-94 cells labelling CRISPR-Cas9 with ab271293 at 1/50 dilution (18.32 ug/ml), followed by ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution (2 ug/ml) (Red). The nuclear counterstain was DAPI (Blue).
Confocal image showing cytoplasmic staining in 293T cells transfected with GFP-tagged Cas9 expression vector.
Secondary antibody only control: Secondary antibody is ab150160 Goat Anti-Rat IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution (2 ug/ml). -
All lanes : Anti-CRISPR-Cas9 antibody [KANI345B] (ab271293) at 1/5000 dilution
Lane 1 : HEK-293 (human embryonic kidney epithelial cell) transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag, whole cell lysate
Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
Lane 4 : C6 (rat glial tumor glial cell), whole cell lysate
Lysates/proteins at 5 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/10000 dilution
Exposure time: 1 secondBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CRISPR-Cas9 with ab271293 at 1/200 (4.58 ug/ml) dilution followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).
Negative control: No staining on the human tonsil. -
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CRISPR-Cas9 with ab271293 at 1/200 (4.58 ug/ml) dilution followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).
Negative control: No staining on the mouse spleen. -
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CRISPR-Cas9 with ab271293 at 1/200 (4.58 ug/ml) dilution followed by a ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Counterstained with Hematoxylin. Heat mediated antigen retrieval using ab93678 (citrate buffer, pH 6.0).
Negative control: No staining on the rat spleen.
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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SDS download
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Datasheet download
References (0)
ab271293 has not yet been referenced specifically in any publications.