Anti-PLD6 Antibody Picoband™

Product Name

Anti-PLD6 Antibody Picoband™

View all Pld6 Antibodies

SKU/Catalog Number

A10904-1

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-PLD6 Antibody Picoband™ catalog # A10904-1. Tested in ELISA, Flow Cytometry, IF, 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-PLD6 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A10904-1)

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 PLD6 recombinant protein (Position: R39-L238).

*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

A10904-1 is reactive to Pld6 in Human

Applications

A10904-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, WB Boster Guarantee

Observed Molecular Weight

28 kDa

Calculated molecular weight

25.041kDa

Background of Pld6

PLD6 (phospholipase D family, member 6) is a protein-coding gene. Among its related super-pathways are choline biosynthesis III and Glycerophospholipid biosynthesis. GO annotations related to this gene include cardiolipin hydrolase activity and protein homodimerization activity. Regulates mitochondrial shape through facilitating mitochondrial fusion. During spermatogenesis, plays a critical role in PIWI-interacting RNA (piRNA) biogenesis (By similarity). piRNAs provide essential protection against the activity of mobile genetic elements. piRNA-mediated transposon silencing is thus critical for maintaining genome stability, in particular in germline cells when transposons are mobilized as a consequence of wide-spread genomic demethylation. Has been shown to be a backbone-non-specific, single strand-specific nuclease, cleaving either RNA or DNA substrates with similar affinity (By similarity). Produces 5' phosphate and 3' hydroxyl termini, suggesting it could directly participate in the processing of primary piRNA transcripts (By similarity). Has been proposed to act as a cardiolipin hydrolase to generate phosphatidic acid at mitochondrial surface. Although it cannot be excluded that it can act as a phospholipase in some circumstances, it should be noted that cardiolipin hydrolase activity is either undetectable in vitro, or very low (PubMed:21397848). In addition, cardiolipin is almost exclusively found on the inner mitochondrial membrane, while PLD6 localizes to the outer mitochondrial membrane, facing the cytosol.

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
Immunofluorescence, 5 µg/ml, Human
Flow Cytometry, 1-3 µg/1x1x10⁶ 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 PLD6 using anti-PLD6 antibody (A10904-1).
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 THP-1 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-PLD6 antigen affinity purified polyclonal antibody (Catalog # A10904-1) 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 PLD6 at approximately 28 kDa. The expected band size for PLD6 is at 28 kDa.

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Figure 2. IHC analysis of PLD6 using anti-PLD6 antibody (A10904-1).
PLD6 was detected in a paraffin-embedded section of human endometrial adenocarcinoma 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-PLD6 Antibody (A10904-1) 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. IHC analysis of PLD6 using anti-PLD6 antibody (A10904-1).
PLD6 was detected in a paraffin-embedded section of human colon adenocarcinoma 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-PLD6 Antibody (A10904-1) 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 4. Flow Cytometry analysis of MCF-7 cells using anti-PLD6 antibody (A10904-1).
Overlay histogram showing MCF-7 cells stained with A10904-1 (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-PLD6 Antibody (A10904-1, 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.

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Figure 5. IF analysis of PLD6 using anti-PLD6 antibody (A10904-1).
PLD6 was detected in a paraffin-embedded section of human oesophagus squama cancer 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 5 μg/mL rabbit anti-PLD6 Antibody (A10904-1) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) was used as secondary antibody at 1:500 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.

Gene/Protein Information For Pld6 (Source: Uniprot.org, NCBI)

Alternative Names

Choline phosphatase 6; EC 3.1.4.-; Mitochondrial phospholipase; MitoPLD; Phosphatidylcholine-hydrolyzing phospholipase D6; phospholipase D family, member 6; Phospholipase D6; PLD 6; Protein zucchini homolog Pld6|4933433K01Rik, Gm10, Zuc, mZuc, mit, mitoPLD|phospholipase D family, member 6|mitochondrial cardiolipin hydrolase|choline phosphatase 6|mitochondrial phospholipase|novel Phospholipase D Active site motif-containing protein|phosphatidylcholine-hydrolyzing phospholipase D6|phospholipase D6|protein zucchini homolog

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