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