Antibodies, Proteins, Kits and Reagents for Life Science

311 587 ₸ Product size

100 µl 311 587 ₸

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Anti-ILKAP antibody [EPR16145] (ab196013)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16145] to ILKAP
Suitable for: WB, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-ILKAP antibody [EPR16145]
See all ILKAP primary antibodies
Description

Rabbit monoclonal [EPR16145] to ILKAP
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Human

WB
Mouse

Rat

Human

See all applications and species data
Immunogen

This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK293T, HAP1, HeLa, 293, Mouse brain and Rat brain lysates.
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.

Application	Species
IP	Human
WB	Mouse Rat Human

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 (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16145
Isotype

IgG
Research areas

Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

All lanes : Anti-ILKAP antibody [EPR16145] (ab196013) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ILKAP knockout HEK293T cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : HAP1 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 43 kDa
Observed band size: 49 kDa
why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab196013 observed at 49 kDa. Red - loading control ab8245 observed at 36 kDa.

ab196013 Anti-ILKAP antibody [EPR16145] was shown to specifically react with ILKAP in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266475 (knockout cell lysate ab258000) was used. Wild-type and ILKAP knockout samples were subjected to SDS-PAGE. ab196013 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 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.
Immunoprecipitation - Anti-ILKAP antibody [EPR16145] (ab196013)

Immunoprecipitation analysis of 293 (Human epithelial cells from embryonic kidney) whole cell extract labeling ILKAP using ab196013 at 1/30 dilution (Lane 2).
Lane 3: IP using Rabbit monoclonal IgG (ab172730) instead of ab196013 in 293 (Human epithelial cells from embryonic kidney) whole cell extract.
Lane 1: Input: 10 μg 293 (Human epithelial cells from embryonic kidney) whole cell extract.
Subsequent WB detection was performed using ab196013 at 1/1000 dilution.
An Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 was used as secondary antibody.
Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ILKAP knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab196013 observed at 43 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab196013 was shown to recognize ILKAP when ILKAP knockout samples were used, along with additional cross-reactive bands. Wild-type and ILKAP knockout samples were subjected to SDS-PAGE. Ab196013 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

Anti-ILKAP antibody [EPR16145] (ab196013) at 1/20000 dilution + HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa
Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

Anti-ILKAP antibody [EPR16145] (ab196013) at 1/20000 dilution + 293 (Human epithelial cells from embryonic kidney) cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa
Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

Anti-ILKAP antibody [EPR16145] (ab196013) at 1/5000 dilution + Mouse brain lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa
Western blot - Anti-ILKAP antibody [EPR16145] (ab196013)

Anti-ILKAP antibody [EPR16145] (ab196013) at 1/1000 dilution + Rat brain lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 43 kDa
Anti-ILKAP antibody [EPR16145] (ab196013)

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