□ 특징

● Cas9 nuclease 검출을 위한 rabbit polyclonal antibody & mouse monoclonal antibody

● Polyclonal Antibody는 Wild-type의 Cas9 뿐만 아니라 변형 Cas9 (Variants)도 인식 가능

● 높은 특이성: mammalian cell lysate에 의한 background가 현저히 낮음

● 고감도: Western blot으로 불과 0.5 ~ 1ng 의 재조합 Cas9 nuclease를 검출할 수 있으며 Cas9 발현 세포 3,500개까지의 signal 인식 가능

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□ 제품설명

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Guide-it™ Cas9Polyclonal Antibody본 제품은 Streptococcuspyogenes유래의 Cas9 단백질 검출을 할 수 있는 rabbit polyclonal antibody이며, 이는 Cas9 variants를 인식 가능하다 (Wild-type Cas9, Cas9 nickase, Cas9 nuclease-deficient mutants). 본 항체를 이용하면 Western blot 방법으로 대부분의 mammalian cell line에서 고감도, 낮은 background로 최소 0.5 ng의 재조합 Cas9을 검출 가능하다. Guide-it™ Cas9 Monoclonal Antibody (Clone TG8C1) 본 제품은 Streptococcus pyogenes 유래의 full-length Cas9 재조합 단백질을 검출할 수 있는 mouseantibody이며 hybridoma cells로부터 생성되었다. 본 항체는 western blot으로 mammalian cell lysate 또는 bacterial celllysate로부터 최소 1ng의 야생형(wilde-type)의 Cas9 단백질을 인식할 수 있다.그림1. The sensitivity of Cas9 polyclonal antibody assessed by Western blot. Various amounts ofrecombinant Cas9 (rCas9) were loaded (Lane 3:0.15 ng, Lane 4: 0.30 ng, Lane 5: 0.625 ng, Lane 6: 1.25 ng, Lane 7: 2.5 ng,Lane 8: 5.0 ng, Lane 9: 10.0 ng, Lane 10: 20.0 ng). Cell lysates prepared from3.5 x 10⁴ cultured cells (Lane 1: HT1080 cells, Lane 2: HEK 293cells) were used as a negative control. The blot was probed with Cas9polyclonal antibody (1:1,000 dilution). For detection, an HRP-conjugated goatanti-rabbit secondary antibody was used (1:5,000 dilution), and signal wasdetected using ECL reagent.그림2. The specificity of Cas9 polyclonal antibody assessed by Western blot. Lysates prepared from various untransfected cell lines (3.5 x 10⁴cultured cells; Lane 1: HepG2, Lane 2: CHO-K1, Lane 3: HeLa, Lane 4: MCF7,Lane5: HEK 293, Lane 6: HT1080, Lane 7: NIH 3T3, Lane 8: no cells) and HEK 293cells expressing Cas9 from the CMV promoter (stable cell line; Lane 10: 2.0 x10³ cells, Lane 11: 5.0 x 10³ cells, Lane 12: 1.0 x 10⁴ cells,Lane 13: 1.5 x 10⁴ cells). One nanogram of recombinant Cas9(rCas) was used as a positive control (Lane 9). The blot was probed with Cas9polyclonal antibody (1:1,000 dilution). For detection, an HRP-conjugated goatanti-rabbit secondary antibody was used (1:5,000 dilution), and signal wasdetected using ECL reagent. 그림 3. Western blot analysisof the Guide-it™ Cas9 Monoclonal Antibody (Clone TG8C1). The Guide-it Cas9Monoclonal Antibody (Clone TG8C1), which was raised in mouse againstrecombinant full-length Cas9 (rCas9), was tested via Western blot analysisusing either increasing concentrations of recombinant Cas9 (Panel A) or lysatesmade from cells transiently expressing Cas9 after plasmid transfection (PanelB). Panel A: Increasing concentrations of rCas9 (1 ng, 2 ng, 3 ng, and 5 ng)were analyzed via Western blotting. Panel B: HEK 293 cells were transientlytransfected with a plasmid encoding Cas9 protein.

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□ 적용

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Western blot detection of Cas9 proteinImmunocytochemistry detection of Cas9 protein

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□ 보존

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-20 ℃

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□ 구성품

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