Antibodies, Proteins, Kits and Reagents for Life Science

338 390 ₸ Product size

100 µl 338 390 ₸

Order now and get it on Tuesday March 02, 2021

Anti-ATAD2 antibody [EPR12730] (ab176319)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR12730] to ATAD2
Suitable for: WB, ICC/IF, Flow Cyt
Knockout validated
Reacts with: Human

Product name

Anti-ATAD2 antibody [EPR12730]
See all ATAD2 primary antibodies
Description

Rabbit monoclonal [EPR12730] to ATAD2
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human ATAD2 aa 50-150 (Cysteine residue). The exact sequence is proprietary.
Database link: Q6PL18
Positive control
- WB: HeLa, MCF-7, T47-D and Saos-2 cell lysates. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
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.

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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR12730
Isotype

IgG
Research areas
- Epigenetics and Nuclear Signaling
- Bromodomains

Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)

Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: ATAD2 knockout HAP1 whole cell lysate (20 µg)

Lanes 1 - 2: Merged signal (red and green). Green - ab176319 observed at 200 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab176319 was shown to specifically react with ATAD2 in wild type cells as signal was lost in ATAD2 knockout cells. Wild-type and ATAD2 knockout samples were subjected to SDS-PAGE. Ab176319 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Flow Cytometry - Anti-ATAD2 antibody [EPR12730] (ab176319)

Flow cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATAD2 (red) with ab176319 at a 1/60 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
Immunocytochemistry/ Immunofluorescence - Anti-ATAD2 antibody [EPR12730] (ab176319)

Immunofluorescent staining of HeLa cells labeling ATAD2 using ab176319 at a 1/100 dilution.
Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)

All lanes : Anti-ATAD2 antibody [EPR12730] (ab176319) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ATAD2 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 159 kDa
Observed band size: 200 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab176319 observed at 200 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.

ab176319 was shown to react with ATAD2 in wild-type HeLa cells in western blot. The band observed in knockout cell line ab264957 (knockout cell lysate ab257359) lane below 200kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HeLa and ATAD2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab176319 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-ATAD2 antibody [EPR12730] (ab176319)

All lanes : Anti-ATAD2 antibody [EPR12730] (ab176319) at 1/1000 dilution

Lane 1 : HeLa lysate
Lane 2 : MCF-7 lysate
Lane 3 : T47-D lysate
Lane 4 : Saos-2 lysate

Lysates/proteins at 10 µg per lane.

Predicted band size: 159 kDa
Anti-ATAD2 antibody [EPR12730] (ab176319)

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