ELISA binding of GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) with recombinant Streptococcus pyogenes CRISPR/Cas9 protein.
Coating antigen: SpCas9, 1 µg/ml.
SpCas9 antibody dilution start from 1000 ng/ml, EC₅₀= 9.17 ng/ml.
Immunofluorescence staining of GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) with SpCas9 in Hela cells.
HeLa cells transfected with PX458 (pSpCas9(BB)-2A-GFP) (green) were fixed with 4% Poly-Formaldehyde (5min) and then blocked in 3% BSA 30min.
The cells were then incubated with GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) at 0.25 μg/ml, 0.5 μg/ml, 1 μg/ml and mouse IgG at 1 μg/ml at room temperature for 2h, followed by a further incubation at 37℃ for 1h with Goat Anti-Mouse IgG Antibody (H&L) [ifluor 555], pAb (GenScript) (red) at 5 μg/ml.
DAPI was used to stain the cell nuclei (blue) at a concentration of 0.2 μg/ml for 2h at room temperature.
Standard curve of SpCas9 Sandwich ELISA. The SpCas9 Sandwich ELISA assay is developed by using GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40) and GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) as capture and detection antibody, respectively. These two antibodies recognize different epitopes.
In this ELISA assay, GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 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 0-10 ng/ml.
Western Blot analysis of HEK293 transfected with various plasmids with GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40). The different HEK293 with transfected with various plasmids indicate the minimum cross reaction of the antibody.
Loading:
Lane 1: 50μg HEK293 cell lysate transfected with SaCas9(BB)-2A-GFP (J7RUA5, Staphylococcus aureus)
Lane 2: 50μg HEK293 cell lysate transfected with StCas9(BB)-2A-GFP (G3ECR1, Streptococcus thermophiles)
Lane 3: 50μg HEK293 cell lysate transfected with FnCpf1(BB)-2A-GFP (A0Q7Q2, Francisella tularensis subsp. novicida (strain U112))
Lane 4: 50μg HEK293 cell lysate transfected with LbCpf1(BB)-2A-GFP (A0A182DWE3, Lachnospiraceae bacterium ND2006)
Lane 5: 50μg HEK293 cell lysate transfected with AsCpf1(BB)-2A-GFP (U2UMQ6, Acidaminococcus sp. (strain BV3L6))
Lane 6: 50μg HEK293 cell lysate transfected with pSpCas9(BB)-2A-GFP (PX458, Q99ZW2, Streptococcus pyogenes serotype M1)
Lane 7: 50μg HEK293 cell lysate (Non-transfected)
Primary Antibody:
Lane 1~7: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 1 μg/ml
Lane 1~7: THEᵀᴹ Anti-α-tubulin mAb (mouse)(1F4) 0.5μg/ml
Secondary antibody:
Goat anti-Mouse IgG (H&L) [IRDye⁸⁰⁰], 0.125 μg/ml
Western Blot of HEK293 transfected with PX458 (pSpCas9(BB)-2A-GFP) or untransfected cell lysates with GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40). The different concentration of antibodies indicates the high affinity and sensitivity of the antibody.
Lane 1: 50 μg HEK293 transfected with PX458 cell Lysate
Lane 2: 50 μg Untransfected HEK293 cell Lysate
Lane 3: 40 ng SpCas9 recombinant protein
Lane 4: 50 μg HEK293 transfected with PX458 cell Lysate
Lane 5: 50 μg Untransfected HEK293 cell Lysate
Lane 6: 40 ng SpCas9 recombinant protein
Lane 7: 50 μg HEK293 transfected with PX458 cell Lysate
Lane 8: 50 μg Untransfected HEK293 cell Lysate
Lane 9: 40 ng SpCas9 recombinant protein
Primary Antibody:
Lane 1~3: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 2 μg/ml
Lane 4~6: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 1 μg/ml
Lane 7~9: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 0.5 μg/ml
Secondary Antibody:
Goat anti-Mouse IgG (H&L) [IRDye⁸⁰⁰] , 0.125 μg/ml
Western Blot of HEK293 transfected with PX458 (pSpCas9(BB)-2A-GFP) or untransfected cell lysates with GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40). The different concentration of cell lysates indicates the high affinity and sensitivity of the antibody.
Lane 1: 50 μg HEK293 transfected with PX458 cell Lysate
Lane 2: 25 μg HEK293 transfected with PX458 cell Lysate
Lane 3: 10 μg HEK293 transfected with PX458 cell Lysate
Lane 4: 50 μg Untransfected HEK293 cell Lysate
Primary Antibody:
GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 1 μg/ml
Secondary Antibody:
Goat anti-Mouse IgG (H&L) [IRDye⁸⁰⁰], 0.125 μg/ml
Western Blot of HEK293 transfected with PX458 (pSpCas9(BB)-2A-GFP) or untransfected cell lysates with two independent antibodies: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) and GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40). The correlated pattern indicates the high specificity of these two antibodies.
Lane 1: 50 μg HEK293 transfected with PX458 cell Lysate
Lane 2: 50 μg Untransfected HEK293 cell Lysate
Lane 3: 40 ng SpCas9 recombinant protein
Lane 4: 50 μg HEK293 transfected with PX458 cell Lysate
Lane 5: 50 μg Untransfected HEK293 cell Lysate
Lane 6: 40 ng SpCas9 recombinant protein
Primary Antibody:
Lane 1~3: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 1 μg/ml
Lane 4~6: GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40) 1 μg/ml
Secondary Antibody:
Goat anti-Mouse IgG (H&L) [IRDye⁸⁰⁰], 0.125 μg/ml
Western Blot of recombinant Streptococcus pyogenes CRISPR/Cas9 protein with two independent antibodies: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) and GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40). The correlated pattern indicates the high specificity of these two antibodies.
Lane 1: 50 ng SpCas9 recombinant protein
Lane 2: 25 ng SpCas9 recombinant protein
Lane 3: 10 ng SpCas9 recombinant protein
Lane 4: 5 ng SpCas9 recombinant protein
Lane 5: 50 ng SpCas9 recombinant protein
Lane 6: 25 ng SpCas9 recombinant protein
Lane 7: 10 ng SpCas9 recombinant protein
Lane 8: 5 ng SpCas9 recombinant protein
Primary Antibody:
Lane 1~4: GenCRISPRᵀᴹ SpCas9 Antibody (4A1) (GenScript, A01935-40) 1 μg/ml
Lane 5~8: GenCRISPRᵀᴹ SpCas9 Antibody (14B6) (GenScript, A01936-40) 1 μg/ml
Secondary Antibody:
Goat anti-Mouse IgG (H&L) [IRDye⁸⁰⁰] , 0.125 μg/ml
GenCRISPR™ SpCas9 Antibody (4A1), mAb, Mouse
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