Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized 293T (Human epithelial cell line from embryonic kidney) cells transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag or Empty vector, labeling CRISPR-Cas9 with ab203933 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

Confocal image showing positive staining on 293T cells transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) 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 : ab203933 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

• ICC

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Immunocytochemistry - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Immunocytochemical analysis of agarose-embedded 293T (Human epithelial cell line from embryonic kidney) cells transfected with Staphylococcus aureus subsp. Aureus Cas9 (pcDNA3.1(+)-Myc-His) labeling CRISPR-Cas9 with ab203933 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Positive staining on 293T cells transfected with Staphylococcus aureus subsp. Aureus Cas9 (pcDNA3.1(+)-Myc-His) is observed.

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

• ICC

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Immunocytochemistry - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Immunocytochemical analysis of agarose-embedded HeLa (Human epithelial cell line from cervix adenocarcinoma) cells transfected with blank pcDNA3.1(+)-Myc-His vector labeling CRISPR-Cas9 with ab203933 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on HeLa cells transfected with blank pcDNA3.1(+)-Myc-His vector.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Immunohistochemical analysis of paraffin-embedded Human endometrium tissue labeling CRISPR-Cas9 with ab203933 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on Human endometrium is observed.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HEK-293 (Human epithelial cell line from embryonic kidney) cells transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag labeling CRISPR-Cas9 with ab203933 at 1/60 dilution (red) compared with a Rabbit IgG,monoclonal [EPR25A] -Isotype control (ab172730) (black). Goat anti Rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

• IP

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Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

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 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag with ab203933 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab203933 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 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag 10μg (Input).

Lane 2 : ab203933 IP in HEK-293 whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag.

Lane 3 : Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab203933 in HEK-293 whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, Staphylococcus aureus subsp. aureus) with Myc-His tag.

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

Exposure time : 1 second.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

All lanes:

Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19799] (<a href='/en-us/products/primary-antibodies/crispr-cas9-antibody-epr19799-ab203933'>ab203933</a>)

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling CRISPR-Cas9 with ab203933 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on rat spleen is observed.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19799] - BSA and Azide free (AB218654)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CRISPR-Cas9 with ab203933 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on mouse spleen is observed.

Counter stained with Hematoxylin.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203933).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.