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Purified anti-CRISPR (CAS9) Antibody

Pricing & Availability
Clone
7A9 (See other available formats)
Regulatory Status
RUO
Other Names
CRISPR associated protein 9, SpyCas9
Isotype
Mouse IgG1, κ
Ave. Rating
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Product Citations
publications
1. 7A9_PUREAF_CRISPR_Antibody_FC_041015
HeLa cells expressing Tet-inducible Flag-tagged S. pyogenes Cas9 were seeded onto glass cover slips and induced with 1.5 µg/ml Doxycyclin for 30 hours. Cells were fixed in 3.7% formaldehyde in PBS, permeablized, and stained with anti-Cas9 (Clone 7A9) antibody at 4°C for 24 hours. After incubation with donkey anti-mouse Alexa Fluor® 488, the nuclei were counter-stained with Hoechst 33342.
  • 1. 7A9_PUREAF_CRISPR_Antibody_FC_041015
    HeLa cells expressing Tet-inducible Flag-tagged S. pyogenes Cas9 were seeded onto glass cover slips and induced with 1.5 µg/ml Doxycyclin for 30 hours. Cells were fixed in 3.7% formaldehyde in PBS, permeablized, and stained with anti-Cas9 (Clone 7A9) antibody at 4°C for 24 hours. After incubation with donkey anti-mouse Alexa Fluor® 488, the nuclei were counter-stained with Hoechst 33342.
  • 2. 7A9_PUREAF_CRISPR_Antibody_WB_041015
    HeLa cells were transiently transfected with Flag-tagged S. pyogenes Cas9 and cells are lysed in hot Laemmli buffer at 30 hours post transfection. Equal amounts of untransfected and transfected cell lysates were separated by 7.5% SDS-PAGE, transferred to nitrocellulose membrane, and incubated at 4°C with the indicated antibodies. Cas9 was used at 0.5 µg/ml.
  • 3. 7A9_PUREAF_CRISPR_Antibody_3_073125
    HAP1 Cas9 Stable Cells (Horizon Cat. No. HD Cas9-011) were fixed and permeabilized with cold methanol and then blocked with 5% FBS. Cells were stained with Purified Mouse IgG1, κ Isotype Control Antibody (Cat. No. 400101) (panel A) or Purified anti-CRISPR (CAS9) (clone 7A9) (panels B and C) followed by Alexa Fluor® 647 Goat anti-Mouse IgG (Cat. No. 405322). Nuclei were counterstained with DAPI (Cat. No. 422801) (overlay in panels A and B). The images were captured on a Revvity Operetta CLS™ High Content Analysis System with a 63X objective. Scale bar: 50 µm
  • 4. 7A9_PUREAF_CRISPR_Antibody_4_073125
    Whole cell extracts (250 µg total protein) prepared from HAP1 Cas9 Stable Cell Line (Horizon Cat No. HD Cas9-011) were immunoprecipitated overnight with 2.0 µg of Purified Mouse IgG1, κ Isotype Control antibody (Cat. No. 401402) or Purified anti-CRISPR (CAS9) antibody (clone 7A9). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with Purified anti-CRISPR (Cas9) antibody (6G12-H11) (Cat. No. 698302). Lane M: Molecular weight marker
  • 5. 7A9_PUREAF_CRISPR_Antibody_5_103025
    Wildtype HAP1 cells (negative control, open histogram) (Horizon Cat. No. C631) or HAP1 Cas9 Stable Cells (positive control, filled histogram) (Horizon Cat. No. HD Cas9-011) were fixed and permeabilized using Cyto-Fast™ Fix/Perm Buffer Set (Cat. No. 426803) and intracellularly stained with Purified anti-CRISPR (CAS9) (clone 7A9) or Purified Mouse IgG1, κ Isotype Control (dashed histogram) (Cat. No. 400101) followed by Alexa Fluor® 647 Goat anti-Mouse IgG (Cat. No. 405322).
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844302 25 µg 72€
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844301 100 µg 203€
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Description

Cas9 is an RNA-guided DNA endonuclease enzyme associated with the CRISPR (Clustered Regularly Interspersed Palindromic Repeats) adaptive immunity system in Streptococcus pyogenes, among other bacteria. Cas9 interrogates and cleaves foreign DNA, such as invading bacteriophage DNA or plasmid DNA. Cas9 performs this interrogation by unwinding foreign DNA and checking if it is complementary to the 20 basepair spacer region of the guide RNA. If the DNA substrate is complementary to the guide RNA, Cas9 cleaves the invading DNA. In this sense, the CRISPR-Cas9 mechanism has a number of parallels with the RNA interference (RNAi) mechanism in eukaryotes.

Product Details
Quality Statement

This product was carefully developed for performance and specificity using the following products from Horizon Discovery, a Revvity company:

HAP1 Cas9 Stable Cell Line (Horizon Cat No. HD Cas9-011)
HAP1 WT (Horizon Cat. No. C631)

Technical Data Sheet (pdf)

Product Details

Verified Reactivity
S. Pyogenes
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Recombinant S. pyogenes Cas9 Protein (N-terminal 250 amino acids) expressed in E. coli.
Formulation
Phosphate-buffered solution, pH 7.2.
Preparation
The antibody was purified by affinity chromatography.
Concentration
1.0 mg/mL
Storage & Handling
This antibody should be handled aseptically as it is free of preservatives such as Sodium Azide. Store this antibody undiluted between 2°C and 8°C. Please note the storage condition for this antibody has been changed from -20°C to between 2°C and 8°C. You can also check the vial label or CoA to find the proper storage conditions.
Application

ICC - Quality tested
WB, IP, ICFC - Verified
Recommended Usage

Each lot of this antibody is quality control tested by immunocytochemistry. For immunocytochemistry, the suggested working dilution is 1.0 - 10 µg/mL. For western blotting, the suggested use of this reagent is 0.5 - 1.0 µg per mL. For flow cytometric staining using our Cyto-Fast™ Fix/Perm Buffer Set, the suggested use of this reagent is  ≤ 0.062 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.

For use of this antibody in intracellular flow cytometry (ICFC), we recommend fixation and permeabilization using Cyto-Fast™ Fix-Perm Buffer set (Cat. No. 426803). Similar results were also obtained using True-Phos™ Perm Buffer (Cat. No. 425401).

Product Citations
  1. Lucas CD, et al. 2022. Sci Immunol. 7:eabm4032. PubMed
  2. Roy IM, et al. 2022. iScience. 25:105171. PubMed
  3. Jo DH, et al. 2023. Mol Ther Nucleic Acids. 31:16. PubMed
  4. Shang P, et al. 2023. Nat Commun. 14:2615. PubMed
  5. Usman WM, et al. 2018. Nat Commun. 9:278. PubMed
  6. Zafra MP, et al. 2018. Nat Biotechnol. 36:888. PubMed
  7. Smeenk L, et al. 2021. Cancer Discov. 11:2868. PubMed
  8. Jang HK, et al. 2021. Sci Adv. 7:. PubMed
  9. Geisinger JM, et al. 2020. Nucleic Acids Res. 48:9067. PubMed
  10. Clements KE, et al. 2020. Nat Commun. 4.706944444. PubMed
  11. Habib O, et al. 2022. Nucleic Acids Res. 50:1187. PubMed
  12. Siadous FA, et al. 2020. Autophagy. :1. PubMed
  13. Scheidmann MC, et al. 2022. Cancer Res. 82:681. PubMed
  14. Freitas GP, et al. 2021. Gene Ther. 28:748. PubMed
  15. Henriksson J, et al. 2019. Cell. 176:882. PubMed
  16. Schleicher EM, et al. 2020. PLoS Genet. 16:e1009176. PubMed
RRID
AB_2749904 (BioLegend Cat. No. 844302)
AB_2565570 (BioLegend Cat. No. 844301)

Antigen Details

Structure
The architectures of Cas9 enzymes define nucleic acid binding clefts to form a central channel where DNA substrates are bound.
Function
The Cas9 protein from S. pyogenes is part of the CRISPR/Cas9 system of acquired immunity that bacteria have evolved to destroy foreign DNA elements, such as bacteriophages.
Biology Area
Cell Biology, Neuroscience
Antigen References

1. Mali P, et al. 2013. Science 339:823.

Gene ID
901176 View all products for this Gene ID
UniProt
View information about CRISPR on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 5    Revision Date: 10-30-2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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