Anti-ILKAP Antibody Picoband™

Product Name

Anti-ILKAP Antibody Picoband™

View all ILKAP/PP2C delta Antibodies

SKU/Catalog Number

A08074-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-ILKAP Antibody Picoband™ catalog # A08074-1. Tested in ELISA, IF, ICC, WB, Flow Cytometry applications. This antibody reacts with Human.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-ILKAP Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A08074-1)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Clone Number

1B9

Isotype

IgG

Immunogen

E.coli-derived human PDK1 recombinant protein (Position: Y114-A436). Human PDK1 shares 94.7% and 95.7% amino acid (aa) sequence identity with mouse and rat PDK1, respectively.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

A08074-1 is reactive to Ilkap in Human

Applications

A08074-1 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

43 kDa

Calculated molecular weight

42.744kDa

Background of ILKAP/PP2C delta

Integrin-linked kinase-associated serine/threonine phosphatase 2C is an enzyme that in humans is encoded by the ILKAP gene. The protein encoded by this gene is a protein serine/threonine phosphatase of the PP2C family. This protein can interact with integrin-linked kinase (ILK/ILK1), a regulator of integrin mediated signaling, and regulate the kinase activity of ILK. Through the interaction with ILK, this protein may selectively affect the signaling process of ILK-mediated glycogen synthase kinase 3 beta (GSK3beta), and thus participate in Wnt signaling pathway.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Human
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of ILKAP using anti-ILKAP antibody (A08074-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human 293T whole cell lysates,
Lane 3: human HepG2 whole cell lysates,
Lane 4: human K562 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-ILKAP antigen affinity purified polyclonal antibody (Catalog # A08074-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for ILKAP at approximately 43 kDa. The expected band size for ILKAP is at 43 kDa.

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Figure 2. IF analysis of ILKAP using anti-ILKAP antibody (A08074-1) and anti-Beta Tubulin antibody (M01857-3).
ILKAP was detected in immunocytochemical section of A549 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-ILKAP Antibody (A08074-1) and mouse anti-Beta Tubulin antibody (M01857-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) and Cy3 Conjugated Goat Anti-Mouse IgG (BA1031) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 2. Flow Cytometry analysis of HepG2 cells using anti-ILKAP antibody (A08074-1).
Overlay histogram showing HepG2 cells stained with A08074-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ILKAP Antibody (A08074-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For Ilkap (Source: Uniprot.org, NCBI)

Alternative Names

DKFZp434J2031; EC 3.1.3.16; FLJ10181; ILKAP; integrin-linked kinase-associated serine/threonine phosphatase 2C; integrin-linked kinase-associated serine/threonine phosphatase; MGC4846; PP2C delta; PP2C-DELTA; protein phosphatase 2c, delta isozyme Ilkap|ILK associated serine/threonine phosphatase|integrin-linked kinase-associated serine/threonine phosphatase 2C|PP2Cdelta|protein phosphatase 2C

For more info on Ilkap, check out the Ilkap Infographic

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In this infographic, you will see the following information for Ilkap: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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