Anti-PDP1/PDP Antibody Picoband™

Product Name

Anti-PDP1/PDP Antibody Picoband™

View all Pyruvate Dehydrogenase Phosphatase/PDP1/PPM2C Antibodies

SKU/Catalog Number

A05607-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-PDP1/PDP Antibody Picoband™ catalog # A05607-2. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.

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-PDP1/PDP Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A05607-2)

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 PDP1/PDP recombinant protein (Position: S37-E537).

*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

A05607-2 is reactive to PDP1 in Human, Mouse, Rat

Applications

A05607-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

53-61 kDa

Calculated molecular weight

61.054kDa

Background of Pyruvate Dehydrogenase Phosphatase/PDP1/PPM2C

Pyruvate dehydrogenase (E1) is one of the three components (E1, E2, and E3) of the large pyruvate dehydrogenase complex. Pyruvate dehydrogenase kinases catalyze phosphorylation of serine residues of E1 to inactivate the E1 component and inhibit the complex. Pyruvate dehydrogenase phosphatases catalyze the dephosphorylation and activation of the E1 component to reverse the effects of pyruvate dehydrogenase kinases. Pyruvate dehydrogenase phosphatase is a heterodimer consisting of catalytic and regulatory subunits. Two catalytic subunits have been reported; one is predominantly expressed in skeletal muscle and another one is is much more abundant in the liver. The catalytic subunit, encoded by this gene, is the former, and belongs to the protein phosphatase 2C (PP2C) superfamily. Along with the pyruvate dehydrogenase complex and pyruvate dehydrogenase kinases, this enzyme is located in the mitochondrial matrix. Mutation in this gene causes pyruvate dehydrogenase phosphatase deficiency. Multiple alternatively spliced transcript variants encoding different isoforms have been identified.

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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 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 PDP1/PDP using anti-PDP1/PDP antibody (A05607-2).
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 PC-3 whole cell lysates,
Lane 2: human U-87MG whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: mouse brain tissue 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-PDP1/PDP antigen affinity purified polyclonal antibody (Catalog # A05607-2) 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 PDP1/PDP at approximately 53-61 kDa. The expected band size for PDP1/PDP is at 61 kDa.

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Figure 2. IF analysis of PDP1/PDP using anti-PDP1/PDP antibody (A05607-2).
PDP1/PDP was detected in an immunocytochemical section of U20S cells. 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-PDP1/PDP Antibody (A05607-2) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Figure 3. Flow Cytometry analysis of U937 cells using anti-PDP1/PDP antibody (A05607-2).
Overlay histogram showing U937 cells stained with A05607-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PDP1/PDP Antibody (A05607-2, 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 PDP1 (Source: Uniprot.org, NCBI)

Alternative Names

[Pyruvate dehydrogenase [acetyl-transferring]]-phosphatase 1, mitochondrial; EC 3.1.3; EC 3.1.3.43; MGC119646; PDH; PDP 1; PDP1; PDPC 1; PDPC; PDPFLJ32517; PPM2C; PPM2CFLJ56179; Protein phosphatase 2C; protein phosphatase 2C, magnesium-dependent, catalytic subunit; pyruvate dehydrogenase (Lipoamide) phosphatase-phosphatase; Pyruvate dehydrogenase phosphatase catalytic subunit 1; Pyruvate Dehydrogenase Phosphatase; pyruvate dehyrogenase phosphatase catalytic subunit 1 PDP1 PDH, PDP, PDPC, PPM2A, PPM2C pyruvate dehydrogenase phosphatase catalytic subunit 1 pyruvate dehyrogenase phosphatase catalytic subunit 1|PDP 1|PDPC 1|[Pyruvate dehydrogenase [acetyl-transferring]]-phosphatase 1, mitochondrial|protein phosphatase 2C, magnesium-dependent, catalytic subunit|protein phosphatase, Mg2+/Mn2+ dependent 2A|pyruvate dehydrogenase (Lipoamide) phosphatase-phosphatase

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