Antibodies, Proteins, Kits and Reagents for Life Science

341 740 ₸ Product size

100 µl 341 740 ₸

Order now and get it on Tuesday March 02, 2021

Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y171] to Chk2 (phospho T68)
Suitable for: IP, WB, Dot blot
Reacts with: Human

Product name

Anti-Chk2 (phospho T68) antibody [Y171]
See all Chk2 primary antibodies
Description

Rabbit monoclonal [Y171] to Chk2 (phospho T68)
Host species

Rabbit
Specificity

The antibody will detect Chk2 phosphorylation on Threonine 68. This antibody may also detect all splice isoforms (2-12) based on sequence homology.
Tested Applications & Species

Application Species
IP
Human

WB
Human

See all applications and species data
Immunogen

Synthetic peptide within Human Chk2. The exact sequence is proprietary.
Positive control
- Cell lysates from HEK-293 cells treated with UV and/or phosphatase.
General notes
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Application	Species
IP	Human
WB	Human

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

Y171
Isotype

IgG
Research areas

Immunoprecipitation - Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

ab32148 (purified) at 1:30 dilution (2µg) immunoprecipitating Chk2 in HEK-293 treated with UV whole cell lysate.
Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) treated with UV whole cell lysate 10µg
Lane 2 (+): ab32148 & HEK-293 treated with UV whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32148 in HEK-293 treated with UV whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
Western blot - Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

All lanes : Anti-Chk2 (phospho T68) antibody [Y171] (ab32148) at 1/1000 dilution (Purified)

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysates
Lane 2 : HEK-293 (Human epithelial cell line from embryonic kidney) treated with UV at 60J/m2 for 2 hours whole cell lysates
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) treated with UV at 60J/m2 for 2 hours whole cell lysates, then the Membrane incubated with phosphatase

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 61 kDa
Observed band size: 70 kDa
why is the actual band size different from the predicted?
Western blot - Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

All lanes : Anti-Chk2 (phospho T68) antibody [Y171] (ab32148) at 1/200 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : Whole cell lysate from HEK-293 (Human epithelial cell line from embryonic kidney) cells, treated with UV at 60J/m2 for 2 hours.
Lane 3 : Whole cell lysate from HEK-293 (Human epithelial cell line from embryonic kidney) cells, treated with UV at 60J/m2 for 2 hours. Membrane incubated with phosphatase.

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 61 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

Blocking/dilution buffer: 5% NFDM/TBST
Immunoprecipitation - Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

ab32148 at 1/20 dilution immunoprecipitating Chk2 (phospho T68) in HEK-293 (human embryonic kidney) whole cell lysate observed at 70kDa (lanes 1 and 2).

Lane 1 (input): HEK-293 cells treated with UV at 60J/m² for 2 hours. Whole cell lysate 10μg

Lane 2 (+): ab32148 + HEK-293 treated with UV at 60J/m² for 2 hours. Whole cell lysate

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32148 in HEK- treated with UV at 60J/m2 for 2 hours. Whole cell lysate

For western blotting, ab32148 at 1/500 dilution was used followed by ab131366 VeriBlot for IP Detection Reagent (HRP) for detection at 1/1000.

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

This image was generated using the unpurified version of the product.
Dot Blot - Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

Dot Blot analysis on immunogen phospho-peptide (A) and non-phospho peptide (B) using anti-Phospho-Chkr2 (pT68) RabMAb (ab32148) dilution 1:500

This image was generated using the unpurified version of the product.
Western blot - Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

All lanes : Anti-Chk2 (phospho T68) antibody [Y171] (ab32148) at 1/1000 dilution

Lane 1 : HCT116 treated with DMSO (control) whole cell lysate
Lane 2 : HCT116 treated with 10 µM etoposide for 24 hours whole cell lysate

Lysates/proteins at 60 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 61 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?

Exposure time: 20 seconds

Non-specific band : 35, 30 kDa

Blocking/Diluting buffer : 5% Milk

This image was generated using the unpurified version of the product.
See Abreview
Anti-Chk2 (phospho T68) antibody [Y171] (ab32148)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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