GenCRISPR™ SaCas9 Antibody (26H10), MAb, Mouse

ELISA binding of GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40) with recombinant Staphylococcus aureus CRISPR/Cas9 protein.
Coating antigen: SaCas9, 1 µg/ml.
SaCas9 antibody dilution start from 1000 ng/ml,EC₅₀= 2.109 ng/ml.

Standard curve of SaCas9 Sandwich ELISA. The SaCas9 Sandwich ELISA assay is developed by using GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40) and GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40) as capture and detection antibody, respectively. These two antibodies recognize different epitopes.
In this ELISA assay, GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40) was labeled with Biotin. GenScript can provide customized conjugation service for this product per customer's request.
The sensitivity is ＜1 ng/ml and the detection range is 0-30 ng/ml.

Western Blot analysis of Hela transfected with various plasmids with GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40). The different Hela transfected with various plasmids indicate the minimum cross reaction of the antibody.
Predicted band size: 124 kDa
Loading:
Lane 1: 50μg Hela cell lysate transfected with StCas9(BB)-2A-GFP (G3ECR1, Streptococcus thermophiles)
Lane 2: 50μg Hela cell lysate transfected with pSpCas9(BB)-2A-GFP (PX458, Q99ZW2, Streptococcus pyogenes serotype M1)
Lane 3: 50μg Hela cell lysate transfected with SaCas9(BB)-2A-GFP (J7RUA5, Staphylococcus aureus)
Lane 4: 50μg Hela cell lysate transfected with AsCpf1(BB)-2A-GFP (U2UMQ6, Acidaminococcus sp. (strain BV3L6))
Lane 5: 50μg Hela cell lysate transfected with FnCpf1(BB)-2A-GFP (A0Q7Q2, Francisella tularensis subsp. novicida (strain U112))
Lane 6: 50μg Hela cell lysate transfected with LbCpf1(BB)-2A-GFP (A0A182DWE3, Lachnospiraceae bacterium ND2006)
Lane 7: 50μg Hela cell lysate (Non-transfected)
Primary Antibody:
Lane 1~7: GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40) 1 μg/ml
Secondary antibody：
Goat anti-Mouse IgG (H&L) [IRDye⁸⁰⁰] , 0.125 μg/ml

Immunofluorescence staining of GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40) with SaCas9 in Hela cells.
HeLa cells transfected with J7RUA5 (SaCas9(BB)-2A-GFP) (colored green) were fixed with 4% Poly-Formaldehyde (5min) and then blocked in 3% BSA 30 min.
The cells were then incubated with GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40) at 0.5 μg/ml mouse IgG at 0.5 μg/ml at room temperature for 2 h, followed by a further incubation at 37°C for 1 h with Goat Anti-Mouse IgG Antibody (H&L) [ifluor 555], pAb (GenScript) (colored red) at 5 μg/ml.
DAPI was used to stain the cell nuclei (colored blue) at a concentration of 0.2 μg/ml for 2 h at room temperature.

Immunoprecipitation of SaCas9 from Hela cell transfected with J7RUA5 (SaCas9(BB)-2A-GFP) by using GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40).
Predicted band size: 124 kDa
Loading:
Lane 1: Hela cell lysate transfected with J7RUA5 (SaCas9(BB)-2A-GFP) 50 μg (Input)
Lane 2: Protein A MagBeads (20 μl) + Purified antibody of A01951-40 (10 μg) + Hela cell lysate transfected with J7RUA5 (SaCas9(BB)-2A-GFP) (200 μg)
Lane 3: Protein A MagBeads (20 μl) + Purified antibody of A01951-40 (10 μg) + Hela cell lysate (200 μg)
Lane 4: Protein A MagBeads (20 μl) + Mouse IgG (10 μg) + Hela cell lysate transfected with J7RUA5 (SaCas9(BB)-2A-GFP) (200 μg)
Lane 5: Protein A MagBeads (20 μl) + Mouse IgG (10 μg) + Hela Cell lysate (200 μg)
Lane 6: Hela Cell lysate (50 μg)
Primary Antibody:
GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40), 1 μg/ml
Secondary antibody：
Goat anti-Mouse IgG (H&L) [IRDye800], 0.125 μg/ml

Western Blot of Hela transfected with J7RUA5 (SaCas9(BB)-2A-GFP) or untransfected cell lysates with GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40). The different concentration of cell lysates indicates the high affinity and sensitivity of the antibody.
Predicted band size: 124 kDa
Predicted band size of recombinant protein: 47.5 kDa
Loading:
Lane 1: 50 μg Hela transfected with SaCas9(BB)-2A-GFP cell lysate
Lane 2: 25 μg Hela transfected with SaCas9(BB)-2A-GFP cell lysate
Lane 3: 10 μg Hela transfected with SaCas9(BB)-2A-GFP cell lysate
Lane 4: 50 μg Untransfected Hela cell lysate
Lane 5: 40 ng SaCas9 recombinant protein
Primary Antibody:
GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40), 1 μg/ml
Secondary Antibody:
Goat anti-Mouse IgG (H&L) [IRDye800], 0.125 μg/ml

Western Blot of Hela transfected with J7RUA5 (SaCas9(BB)-2A-GFP) or untransfected cell lysates with two independent antibodies: GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40) and GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40). The correlated pattern indicates the high specificity of these two antibodies.
Predicted band size: 124 kDa
Predicted band size of recombinant protein: 47.5 kDa
Loading:
Lane 1: 50 μg Hela transfected with SaCas9(BB)-2A-GFP cell lysate
Lane 2: 50 μg Untransfected Hela cell lysate
Lane 3: 40 ng SaCas9 recombinant protein
Lane 4: 50 μg Hela transfected with SaCas9(BB)-2A-GFP cell lysate
Lane 5: 50 μg Untransfected Hela cell lysate
Lane 6: 40 ng SaCas9 recombinant protein
Primary Antibody:
Lane 1~3: GenCRISPRᵀᴹ SaCas9 Antibody (11C12) (GenScript, A01951-40), 1 μg/ml
Lane 4~6: GenCRISPRᵀᴹ SaCas9 Antibody (26H10) (GenScript, A01952-40), 1 μg/ml
Secondary Antibody:
Goat anti-Mouse IgG (H&L) [IRDye800] , 0.125 μg/ml

GenCRISPR™ SaCas9 Antibody (26H10), mAb, Mouse

The product is specific for Staphylococcus aureus CRISPR/Cas9. This antibody binds with recombinant Staphylococcus aureus CRISPR/Cas9 protein in ELISA and Endogenous overexpressed Staphylococcus aureus CRISPR/Cas9 in immunofluorescence, western blot.

产品编号	A01952
规格	40 μg 大单询价
数量
价格	￥990


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产品概述

Specificity	The product is specific for Staphylococcus aureus CRISPR/Cas9. This antibody binds with recombinant Staphylococcus aureus CRISPR/Cas9 protein in ELISA and Endogenous overexpressed Staphylococcus aureus CRISPR/Cas9 in immunofluorescence, western blot.
Host Species	Mouse
Immunogen	Recombinant Staphylococcus aureus CRISPR/Cas9

产品属性

Concentration	0.5 mg/ml, lyophilized with PBS, pH 7.4, contains 0.02% sodium azide. (Might be eligible for customized bulk orders.)
Reconstitution	Reconstitute the lyophilized antibody with deionized water (or equivalent) to a final concentration of 0.5 mg/ml.
Purification	Protein A affinity column
Clone ID	26H10

图例

技术背景

Target Background

Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) protein 9 system provides a robust and multiplexable genome editing tool, enabling researchers to precisely manipulate specific genomic elements, and facilitating the elucidation of target gene function in biology and diseases. CRISPR/Cas9 genome editing allows for double-stranded DNA breaks at specific sequences to efficiently disrupt, excise, mutate, insert, or replace genes. The best characterized CRISPR-associated nucleases are the Cas9 proteins from Streptococcus pyogenes and Staphylococcus aureus. It is important that the precision of transfection and the level of Cas9 expression should be controlled strictly during the editing processes by using specific anti-CRISPR/Cas9 antibodies.

Synonyms

Mouse monoclonal to Staphylococcus aureus Cas9/SaCas9

GenCRISPR™ SaCas9 Antibody (26H10), mAb, Mouse

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