Anti-Pleckstrin/PLEK Antibody Picoband™

Product Name

Anti-Pleckstrin/PLEK Antibody Picoband™

View all Pleckstrin Antibodies

SKU/Catalog Number

A06656-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Pleckstrin/PLEK Antibody Picoband™ catalog # A06656-3. Tested in ELISA, Flow Cytometry, IHC, WB 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-Pleckstrin/PLEK Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A06656-3)

Host

Rabbit

Contents

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

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived human Pleckstrin/PLEK recombinant protein (Position: M47-K308).

*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

A06656-3 is reactive to PLEK in Human

Applications

A06656-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

40 kDa

Calculated molecular weight

40.125kDa

Background of Pleckstrin

Enables phosphatidylinositol-3,4-bisphosphate binding activity; protein homodimerization activity; and protein kinase C binding activity. Involved in several processes, including G protein-coupled receptor signaling pathway; actin cytoskeleton organization; and positive regulation of supramolecular fiber organization. Located in cytoplasm and ruffle membrane.

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
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5 μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of Pleckstrin/PLEK using anti-Pleckstrin/PLEK antibody (A06656-3).
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 HEL whole cell lysates,
Lane 2: human U937 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-Pleckstrin/PLEK antigen affinity purified polyclonal antibody (Catalog # A06656-3) 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 Pleckstrin/PLEK at approximately 40 kDa. The expected band size for Pleckstrin/PLEK is at 40 kDa.

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Figure 2. IHC analysis of Pleckstrin/PLEK using anti-Pleckstrin/PLEK antibody (A06656-3).
Pleckstrin/PLEK was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Pleckstrin/PLEK Antibody (A06656-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. Flow Cytometry analysis of HL-60 cells using anti-Pleckstrin/PLEK antibody (A06656-3).
Overlay histogram showing HL-60 cells stained with A06656-3 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Pleckstrin/PLEK Antibody (A06656-3, 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 PLEK (Source: Uniprot.org, NCBI)

Alternative Names

FLJ27168; p47; P47Platelet 47 kDa protein; pleckstrin PLEK P47 pleckstrin pleckstrin|platelet 47 kDa protein

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