Anti-CRISPR-Cas9 antibody [EPR19619-18]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded 293T cells transfected with Streptococcus Thermophilus Cas9 (pcDNA3.1(+)-Myc-His) with ab202637 at 1/100 dilution. Goat Anti-Rabbit IgG H&L (HRP), ab97051 at 1/500 dillution was used as the secondary antibody. Counterstained with Hematoxylin. Secondary only negative control also shown. Heat mediated antigen retrieval using Tris/EDTA Buffer, pH 9.0 was performed.

Positive staining on the 293T cells transfected with Streptococcus Thermophilus Cas9.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

Immunohistochemical analysis of formalin-fixed, paraffin-embedded 293T cells transfected blank pcDNA3.1(+)-Myc-His vector with ab202637 at 1/100 dilution. Goat Anti-Rabbit IgG H&L (HRP), ab97051 at 1/500 dillution was used as the secondary antibody. Counterstained with Hematoxylin. Secondary only negative control also shown. Heat mediated antigen retrieval using Tris/EDTA Buffer, pH 9.0 was performed.

Negative on the 293T cells transfected blank pcDNA3.1(+)-Myc-His vector.

• IP

Supplier Data

Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

CRISPR-Cas9 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate with ab202637 at 1/30 dilution.

Western blot was performed from the immunoprecipitate using ab202637 at 1/1000 dilution.

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

Lane 1 : 293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate, 10μg (Input).

Lane 2 : ab202637 IP in 293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate.

Lane 3 : Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730) instead of ab202637 in 293T cells transfected with Streptococcus thermophilus CRISPR-Cas9-myc whole cell lysate.

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637)

false

• WB

Supplier Data

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1-2 : 5 seconds; Lane 3 : 3 minutes.

All lanes:

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

Lane 1:

Untransfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (control) at 20 µg

Lane 2:

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

Lane 3:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A) with 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 1/100000 dilution

Predicted band size: 164 kDa

false

• WB

Supplier Data

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

Blocking/Dilution buffer : 5% NFDM/TBST.

The immunogen is located at the N-terminus.

All lanes:

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

Lane 1:

Untransfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate (control) 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-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-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-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 1/100000 dilution

Predicted band size: 164 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

Lane 1:

Untransfected HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate (control) at 20 µg

Lane 2:

HEK-293 whole cell lysate transfected with CRISPR-Cas9 (A1IQ68, Neisseria meningitidis serogroup A) with His-tag at 20 µg

Lane 3:

HEK-293 whole cell lysate transfected with CRISPR-Cas9 (Q03JI6, Streptococcus thermophilus (strain ATCC BAA-491 / LMD-9)) with 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 1/100000 dilution

Predicted band size: 164 kDa

false

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (AB202637)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-CRISPR-Cas9 antibody [EPR19619-18] (ab202637) at 1/5000 dilution

Lane 1:

HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus, N–terminal) with GFP-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

false

Exposure time: 10s