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CRISPR/Cas9 Monoclonal Antibody [7A9-3A3]-20ul
Catalogue Number: IQ651S
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Datasheet
| Primary Description: | Mouse Anti- CRISPR/Cas9 [7A9-3A3] |
| Target Antigen: | CRISPR/Cas9 |
| Catalogue Number: | IQ651S |
| Quantity: | 0.02mg |
| Concentration: | 1mg/ml |
| Clone: | 7A9-3A3 |
| Host: | Mouse |
| Isotype: | IgG1k |
| Myeloma/Fusion Partners: | X63.Ag8-653 |
| Immunogen: | This antibody was raised against a recombinant protein within the N-terminal region of Streptococcus pyogene Cas9 |
| Species Reactivity: | Streptococcus pyogene |
| Purification: | Protein A |
| Format: | Purified antibody (from supernatant) containing PBS + 0.09% sodium azide |
| Applications: | IB, IP, ICC/IF, WB |
| Dilutions: | Optimal antibody dilution should be determined by titration, however as a guideline try at;IB 0.1-1µg/ml |
| Also Known As: | Cas9 antibodyCRISPR-associated endonuclease Cas9/Csn1 antibodyCRISPR-Cas9/Csn1 antibodycsn1 antibodySpyCas9 antibody |
| Pubmed ID(s): | 33398349, 33441761, 33767386, 34035409, 34321211, 34430300, 32550896, 32661087, 32986839, 32602833, 31964705, 32267915, 31980609, 31900423, 31051099, 30805613, 31361218, 31410472, 30858603, 30617347, 31184714, 31006810, 30073181, 30082871, 30291237, 30213867, 29519936, 29759820, 30537986, 29927077, 27820801, 27486969, 28694259, 28624187, 29141659, 28851710, 27499201, 27638686, 26245833 |
| Entrez Gene ID(s): | 901176 |
| SwissProt ID(s): | q99zw2 |
Citations
| Citation Count: | 39 |
| Citations: | Li K et al. Interrogation of enhancer function by enhancer-targeting CRISPR epigenetic editing. Nat Commun 11:485 (2020). Maggio I et al. Integrating gene delivery and gene-editing technologies by adenoviral vector transfer of optimized CRISPR-Cas9 components. Gene Ther 27:209-225 (2020). Maji B et al. A High-Throughput Platform to Identify Small-Molecule Inhibitors of CRISPR-Cas9. Cell 177:1067-1079.e19 (2019). Qiu W et al. Determination of local chromatin interactions using a combined CRISPR and peroxidase APEX2 system. Nucleic Acids Res 47:e52 (2019). Zuo Z et al. Structural and functional insights into the bona fide catalytic state of Streptococcus pyogenes Cas9 HNH nuclease domain. Elife 8:N/A (2019). Josipovic G et al. Antagonistic and synergistic epigenetic modulation using orthologous CRISPR/dCas9-based modular system. Nucleic Acids Res 47:9637-9657 (2019). Park H et al. In vivo neuronal gene editing via CRISPR-Cas9 amphiphilic nanocomplexes alleviates deficits in mouse models of Alzheimer's disease. Nat Neurosci 22:524-528 (2019). Peters JM et al. Enabling genetic analysis of diverse bacteria with Mobile-CRISPRi. Nat Microbiol 4:244-250 (2019). Korsholm LM et al. Double-strand breaks in ribosomal RNA genes activate a distinct signaling and chromatin response to facilitate nucleolar restructuring and repair. Nucleic Acids Res 47:8019-8035 (2019). Kounatidou E et al. A novel CRISPR-engineered prostate cancer cell line defines the AR-V transcriptome and identifies PARP inhibitor sensitivities. Nucleic Acids Res 47:5634-5647 (2019). Simhadri VL et al. Prevalence of Pre-existing Antibodies to CRISPR-Associated Nuclease Cas9 in the USA Population. Mol Ther Methods Clin Dev 10:105-112 (2018). Vakulskas CA et al. A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells. Nat Med 24:1216-1224 (2018). Xu C et al. Targeting of NLRP3 inflammasome with gene editing for the amelioration of inflammatory diseases. Nat Commun 9:4092 (2018). Huynh N et al. A Drosophila CRISPR/Cas9 Toolkit for Conditionally Manipulating Gene Expression in the Prothoracic Gland as a Test Case for Polytene Tissues. G3 (Bethesda) 8:3593-3605 (2018). Katigbak A et al. Inducible Genome Editing with Conditional CRISPR/Cas9 Mice. G3 (Bethesda) 8:1627-1635 (2018). Tsai YT et al. Clustered Regularly Interspaced Short Palindromic Repeats-Based Genome Surgery for the Treatment of Autosomal Dominant Retinitis Pigmentosa. Ophthalmology 125:1421-1430 (2018). Chen N et al. A novel FLI1 exonic circular RNA promotes metastasis in breast cancer by coordinately regulating TET1 and DNMT1. Genome Biol 19:218 (2018). Liu X et al. CAPTURE: In Situ Analysis of Chromatin Composition of Endogenous Genomic Loci by Biotinylated dCas9. Curr Protoc Mol Biol 123:e64 (2018). Maji B et al. Multidimensional chemical control of CRISPR-Cas9. Nat Chem Biol 13:9-11 (2017). WB ; Human . Zhang Y et al. CRISPR Cas9-guided chromatin immunoprecipitation identifies miR483 as an epigenetic modulator of IGF2 imprinting in tumors. Oncotarget 8:34177-34190 (2017). Aslan Y et al. High-efficiency non-mosaic CRISPR-mediated knock-in and indel mutation in F0 Xenopus. Development 144:2852-2858 (2017). Lattanzi A et al. Correction of the Exon 2 Duplication in DMD Myoblasts by a Single CRISPR/Cas9 System. Mol Ther Nucleic Acids 7:11-19 (2017). ICC/IF, WB ; Human . Kim S et al. Rescue of high-specificity Cas9 variants using sgRNAs with matched 5' nucleotides. Genome Biol 18:218 (2017). Mellott DO et al. Notch signaling patterns neurogenic ectoderm and regulates the asymmetric division of neural progenitors in sea urchin embryos. Development 144:3602-3611 (2017). Howden SE et al. A Cas9 Variant for Efficient Generation of Indel-Free Knockin or Gene-Corrected Human Pluripotent Stem Cells. Stem Cell Reports 7:508-17 (2016). WB ; Human . Lin CC & Potter CJ Non-Mendelian Dominant Maternal Effects Caused by CRISPR/Cas9 Transgenic Components in Drosophila melanogaster. G3 (Bethesda) N/A:N/A (2016). IHC - Wholemount . Lin S et al. In Vivo Transcriptional Activation Using CRISPR/Cas9 in Drosophila. Genetics 201:433-42 (2015). |
Protocols
Immunofluorescence protocol - Formaldehyde fixation
- Collect cells from T.c.unit and remove media from petri dish using suction.
- Wash with 1x PBS and remove.
- Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperature on an orbital shaker.
- Remove PFA and incubate in 0.5% Triton X-IOO in 1x PBS for 5 minutes at room temperature.
- Prepare blocking reagent, this is also the antibody diluent.
- Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker.
- Block with 1 % NCS and 1x PBS for 30 minutes at room temperature.
- Prepare primary antibodies (50?l/coverslip) and moist staining chambers.
- Wash cells 2x with lx PBS at room temperature and air dry briefly.
- Incubate with primary antibody for 1 hr at room temperature in the dark in staining chambers. During this time prepare the secondary antibody.
- Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
- Incubate with secondary antibody for 1 hour at room temperature in the dark in staining chambers.
- Wash cells 5x with 1x PBS.
- Mount in Dapi.
Solutions (prepare fresh the same day of staining).
- * 1x Phosphate buffered saline.
- * Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS).
- * Fixation solution: 3.5% Para formaldehyde.
1.75g PFA in 20 ml d.H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C until dissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7.4. Complete volume with d.H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips.
Immunofluorescence protocol - Methanol/acetone fixation
- Collect cells from T.C.unit and remove media from petri dish using suction.
- Wash with 1x PBS and remove.
- Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice.
- Prepare blocking reagent, this is also the diluent for the antibodies.
- Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker.
- Block with 1% NCS and Ix PBS for 30 minutes at RT.
- Prepare primary antibodies (50?l/coverslip) and moist staining chambers.
- Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes.
- Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During this time prepare secondary antibody.
- Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)
- Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers.
- Wash cells 5x with 1x PBS.
- Mount in Dapi.
Solutions (prepare fresh the same day of staining)
- * 1x Phosphate buffered saline.
- * Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS).
- * Fixation solution: methanol:acetone 1: 1 ice cold.
Western Blotting Protocol
- Transfer gel to PDVF or nitrocellulose membrane
- Place membrane in plastic tray in blocking buffer for one hour with agitation
- Rinse in wash buffer
- Incubate in wash buffer plus primary antibody for one hour
- Wash 6 X 5 minutes with wash buffer
- Incubate in wash buffer plus secondary antibody for one hour
- Wash 6X 5 minutes with wash buffer
- Detect (e.g. ECL, Amersham according to manufacturers instructions)
Wash buffer
PBS + 0.1% Tween 20
Blocking buffer
Wash buffer + 5% dried milk powder
The concentration of antibodies used depends on each antibody, the amount of antigen and the detection method used. Generally, dilution is in the range of a few hundred times dilution to a few thousand times dilution, but usually has to be determined empirically.
FAQ's
How do I place an order?
When placing an order we require a purchase order number, plus name and contact details of the purchaser, and the person who will be using the product. We will add VAT for every order received from an EU country, unless you are VAT exempt. For such VAT exempt customers we will need to receive a copy of the exemption certificate with the order.
Orders can be placed either by our website, via email or by mail. Please see our contact us page for more details
All orders are subject to availability. Prices of products do not include shipping, VAT or import duties, where these are applicable.
Price and other information provided are subject to change without notice. While every effort is made to keep information provided Up to date, ImmuQuest will not be liable if errors should occur in such information.
Formal acceptance of an order will take place when the goods are dispatched. If prices should be changed between the time of receipt of an order and dispatch, ImmuQuest will contact you in advance.
Ordering via the website
To order via the website you will need to find the product(s) you are looking for by either using our search, browsing our categories or using our full antibody catalogue.
Once you have found the antibodies you are looking for you can view more detailed information about a specific antibody by clicking the product or the 'View' button. Once you are sure it is the antibody you are looking for you can add it to your order by clicking the 'Add To Order' button.
This places the antibody in your order basket, you can add as many antibodies as you wish. Once you have selected all of the antibodies you wish to order then click the checkout button. If you are not logged in you will be asked to login or to register, if you are logged in you will be taken to the checkout page.
On the checkout page you are able to double check your order and enter a purchase order number if you have one.
Once you are happy your order is correct you can purchase the order with debit/credit card details and your order will be submitted. Once we have received and checked your order we will contact you to arrange payment and delivery.
How long does delivery take?
Delivery usually takes no more than one week from reciept of an order.
Do you guarantee your antibodies?
Yes. Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.
If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.
Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.
Do you charge VAT?
If you are in the EU and you are no exempt from paying VAT then we must charge VAT (charged at the prevailing UK rate).
Customers that are exempt need to fax an exemption form showing their vat exemption number.
What if I don't have a purchase order number?
You can still place an order without providing a purchase order number but we must recieve payment before your order is despatched.
Non-website orders
Non-website orders can be placed directly with Immuquest at sales@immuquest.com or click on the "contact us" tab for address details
Delivery
Offline Orders
For offline orders please call +44 (0) 1642 713533 or click here to email us.
Delivery & returns
We use federal Express for UK & international shipments, andfor standard shipments the cost is £30 UK & £55 international. If the customer uses their account, we charge a £20 handling fee (UK & international)
We are strongly committed to providing the best quality products, and the best service possible.
Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.
If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.
Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.
Product Guarantee
We are strongly committed to providing the best quality products, and the best service possible. In order to do this we depend on your feedback.
Should any product not perform as described in the product literature, it will be replaced or a full refund will be given. The customer must notify ImmuQuest within 30 days after the goods have been received to request a replacement, forwarding complete test data as requested by ImmuQuest.
If items are ordered incorrectly by the customer, ImmuQuest will consider taking them back as long as they have been stored correctly and have not been opened or tampered with. Such orders may be subject to a 15% restocking charge on the items plus any shipping costs.
Requests for returns must have prior authorization from ImmuQuest, and must be made within 7 days of receipt of the items. Items must be returned in the same or equivalent packaging as originally dispatched, and by an equivalent method of delivery.
If you have any questions please contact us using the following details:
ImmuQuest Ltd
Springboard
Stokesley Business Park
24 Ellerbeck Way
Stokesley
TS9 5JZ
UK
Tel: 01642 713533
Fax: 01642 713988
International Tel: +44 1642 713533
International Fax: +44 1642 713988
When placing an order ImmuQuest require a purchase order number, plus name and contact details of the purchaser, and the person who will be using the product. ImmuQuest will also need a VAT number for customers in the European Union.UK customers that are VAT exempt need to fax an exemption certificate.
Orders can be placed either by our website, via email or by mail. Please see our contact us page for more details. All orders are subject to availability. Prices of products do not include shipping, VAT or import duties where these are applicable.
Price and other information provided are subject to change without notice. While every effort is made to keep information provided Up to date, ImmuQuest will not be liable if errors should occur in such information. Formal acceptance of an order will take place when the goods are dispatched. If prices should be changed between the time of receipt of an order and dispatch, ImmuQuest will contact you in advance.
