Anti-CRISPR-Cas9 antibody [EPR19633]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19633] (AB202657)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag or Empty vector, labeling CRISPR-Cas9 with ab202657 at 1/100 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150079) secondary antibody at 1/1000 dilution.

Confocal image showing Positive staining on HeLa cells transfected with CRISPR-Cas9(G3ECR1, Streptococcus thermophilus) with Myc-His tag.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab202657 at 1/100 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CRISPR-Cas9 antibody [EPR19633] (AB202657)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293 (Human epithelial cell line from embryonic kidney) cells transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag labeling CRISPR-Cas9 with ab202657 at 1/60 dilution (red) compared with a RabbitIgG,monoclonal [EPR25A]-Isotype control (ab172730) (black). Goat anti Rabbit IgG (Alexa Fluorr^® 488) at 1/2000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19633] (AB202657)

CRISPR-Cas9 was immunoprecipitated from 0.35 mg of HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag with ab202657 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab202657 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : HEK-293 whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag 10μg (Input).

Lane 2 : ab202657 IP in HEK-293 whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag.

Lane 3 : Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab202657 in HEK-293 whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19633] (ab202657)

false

• WB

Supplier Data

Western blot - Anti-CRISPR-Cas9 antibody [EPR19633] (AB202657)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CRISPR-Cas9 antibody [EPR19633] (ab202657) at 1/20000 dilution

Lane 1:

Empty vector with GFP-Myc tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 2:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag at 20 µg

Lane 3:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, N–terminal aa1-800) with GFP-Myc tag at 20 µg

Lane 4:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, C–terminal aa801-1409) with GFP-Myc tag at 20 µg

Lane 5:

Empty vector with Myc-His tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg

Lane 6:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A / serotype 4A (strain Z2491)) with Myc-His tag at 20 µg

Lane 7:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag at 20 µg

Lane 8:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (Q03JI6, Streptococcus thermophilus (strain ATCC BAA-491 / LMD-9)) with Myc-His tag at 20 µg

Lane 9:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 100000 Cells

Predicted band size: 164 kDa

false

Exposure time: 5s

• WB

Unknown

Western blot - Anti-CRISPR-Cas9 antibody [EPR19633] (AB202657)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CRISPR-Cas9 antibody [EPR19633] (ab202657) at 1/20000 dilution

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, C–terminal aa801-1409) with GFP-Myc tag at 20 µg

Lane 2:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg

Lane 4:

Rat embryo lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 164 kDa

Observed band size: 86 kDa

false

Exposure time: 5s