Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸

Order now and get it on Wednesday March 10, 2021

Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19799] to CRISPR-Cas9
Suitable for: WB, ICC/IF, Flow Cyt, IP, IHC-P, ICC
Reacts with: Staphylococcus aureus

Product name

Anti-CRISPR-Cas9 antibody [EPR19799]
See all CRISPR-Cas9 primary antibodies
Description

Rabbit monoclonal [EPR19799] to CRISPR-Cas9
Host species

Rabbit
Tested applications

Suitable for: WB, ICC/IF, Flow Cyt, IP, IHC-P, ICCmore details
Species reactivity

Reacts with: Staphylococcus aureus
Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293 whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, S.aureus) with Myc-His tag. ICC: HeLa cells transfected with S.aureus Cas9 (pcDNA3.1(+)-Myc-His). ICC/IF: 293T cells transfected with CRISPR-Cas9 (J7RUA5, S.aureus) with Myc-His tag. Flow Cyt: HEK-293 whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, S.aureus) with Myc-His tag. IP: HEK-293 whole cell lysate transfected with CRISPR-Cas9 (J7RUA5, S.aureus) with Myc-His tag.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR19799
Isotype

IgG

Western blot - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

All lanes : Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933) 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
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
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
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
Lane 5 : Empty vector with Myc-His tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
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
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
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
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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 124 kDa
Observed band size: 124 kDa

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.
Western blot - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

All lanes : Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933) at 1/20000 dilution

Lane 1 : 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
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate
Lane 4 : Rat embryo lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 124 kDa
Observed band size: 124 kDa

Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.
Flow Cytometry - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.
Immunoprecipitation - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-CRISPR-Cas9 antibody [EPR19799] (ab203933)

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