Antibodies, Proteins, Kits and Reagents for Life Science

314 937 ₸ Product size

100 µl 314 937 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR18991] to CRISPR-Cas9
Suitable for: ICC, IHC-P, ICC/IF, WB, Flow Cyt
Reacts with: Streptococcus pyogenes

Product name

Anti-CRISPR-Cas9 antibody [EPR18991]
See all CRISPR-Cas9 primary antibodies
Description

Rabbit monoclonal [EPR18991] to CRISPR-Cas9
Host species

Rabbit
Tested applications

Suitable for: ICC, IHC-P, ICC/IF, WB, Flow Cytmore details
Species reactivity

Reacts with: Streptococcus pyogenes
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB and Flow Cyt: HEK-293 whole cell lysate transfected with CRISPR-Cas9 (Q99ZW2, Streptococcus pyogenes serotype M1) with GFP-Myc tag. IHC: 293T cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc). ICC/IF: 293T cells transfected with CRISPR-Cas9 with GFP-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.

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

Immunogen affinity purified
Clonality

Monoclonal
Clone number

EPR18991
Isotype

IgG

Western blot - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

All lanes : Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380) at 1/20000 dilution

Lane 1 : 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 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: 158 kDa
Observed band size: 184 kDa
why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

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-GFP or GFP only, labeling CRISPR-Cas9 with ab189380 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 647) (ab150079) secondary antibody at 1/1000 dilution.

Confocal image showing positive staining on 293T cells transfected with CRISPR-Cas9 with GFP-tag.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows:-

-ve control 1: ab189380 at 1/500 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.

-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150079 (Alexa Fluor^®647 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on rat cerebrum is observed.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

Flow cytometry analysis of 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) labelling CRISPR-Cas9 (red) with ab189380 at dilution of 1/70. The secondary antibody used was Alexa Fluor^® 488 goat-anti-rabbit IgG (1/2000). Cells were fixed with 4% paraformaldehyde. Isotype control antibody was (ab172730) Rabbit monoclonal IgG (black).
Immunocytochemistry - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc) labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Positive staining on 293T cells transfected with Streptococcus pyogenes serotype M1 Cas9 (pcDNA3.1(+)-GFP-Myc) is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Western blot - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

All lanes : Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380) 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 : Empty vector with Myc-His tag (vector control) transfected HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 5 : 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 6 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with CRISPR-Cas9 (G3ECR1, Streptococcus thermophilus) with Myc-His tag
Lane 7 : 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 8 : 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: 158 kDa
Observed band size: 184 kDa why is the actual band size different from the predicted?

Exposure time: 3 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

Immunocytochemical analysis of agarose-embedded sections of 293T (Human epithelial cell line from embryonic kidney) cells transfected with blank pcDNA3.1(+)-GFP-Myc vector labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

Counter stained with Hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CRISPR-Cas9 antibody [EPR18991] (ab189380)

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on Human stomach 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 [EPR18991] (ab189380)

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labeling CRISPR-Cas9 with ab189380 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

No staining on mouse stomach 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 [EPR18991] (ab189380)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com