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SKU:PA2131
Pack Size:100μg/vial
Validated Species:Human, Mouse, Rat
Conjugate:No
Clonality:Polyclonal
Host:Rabbit
Application:Flow Cytometry, IHC-P, IHC-F, ICC, WB
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Overview

Product Name Anti-PLK2/Snk Antibody
SKU/Catalog Number PA2131
Description Rabbit IgG polyclonal antibody for Serine/threonine-protein kinase PLK2(PLK2) detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human;Mouse;Rat.
Cite This Product Anti-PLK2/Snk Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA2131)
Replacement Item This antibody may replace the following items: sc-32250|sc-32246|sc-34843|sc-34842|sc-20731|sc-373983|sc-34840|sc-1410|sc-373984|sc-365826|sc-101501|sc-514019 from Santa Cruz Biotechnology.
Host Rabbit
Isotype N/A
Validated Species Human, Mouse, Rat
Application Flow Cytometry, IHC-P, IHC-F, ICC, WB

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Recommended Detection Systems Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
Immunogen A synthetic peptide corresponding to a sequence at the C-terminus of human PLK2(666-685aa SSELKNRMEYALNMLLQRCN), different from the related mouse sequence by one amino acid, and from the related rat sequence by two amino acids.
Cross Reactivity No cross reactivity with other proteins
Pack Size 100μg/vial

Properties

Clonality Polyclonal
Form Lyophilized
Contents Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
*carrier free antibody available upon request.
Concentration Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Storage At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time.Avoid repeated freezing and thawing.
Purification Immunogen affinity purified.
Isotype N/A

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name PLK2
Protein Name Serine/threonine-protein kinase PLK2
Molecular Weight 78237 MW
Protein Function Tumor suppressor serine/threonine-protein kinase involved in synaptic plasticity, centriole duplication and G1/S phase transition. Polo-like kinases act by binding and phosphorylating proteins are that already phosphorylated on a specific motif recognized by the POLO box domains. Phosphorylates CENPJ, NPM1, RAPGEF2, RASGRF1, SNCA, SIPA1L1 and SYNGAP1. Plays a key role in synaptic plasticity and memory by regulating the Ras and Rap protein signaling: required for overactivity-dependent spine remodeling by phosphorylating the Ras activator RASGRF1 and the Rap inhibitor SIPA1L1 leading to their degradation by the proteasome. Conversely, phosphorylates the Rap activator RAPGEF2 and the Ras inhibitor SYNGAP1, promoting their activity. Also regulates synaptic plasticity independently of kinase activity, via its interaction with NSF that disrupts the interaction between NSF and the GRIA2 subunit of AMPARs, leading to a rapid rundown of AMPAR-mediated current that occludes long term depression. Required for procentriole formation and centriole duplication by phosphorylating CENPJ and NPM1, respectively. Its induction by p53/TP53 suggests that it may participate in the mitotic checkpoint following stress. .
Tissue Specificity Expressed at higher level in the fetal lung, kidney, spleen and heart. .
Sequence Similarities Belongs to the protein kinase superfamily. Ser/Thr protein kinase family. CDC5/Polo subfamily.
Subcellular Localization Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, centriole . Cell projection, dendrite . Localizes to centrosomes during early G1 phase where it only associates to the mother centriole and then distributes equally to both mother and daughter centrioles at the onset of S phase.
Uniprot ID Q9NYY3
Alternative Names Serine/threonine-protein kinase PLK2;2.7.11.21;Polo-like kinase 2;PLK-2;hPlk2;Serine/threonine-protein kinase SNK;hSNK;Serum-inducible kinase;PLK2;SNK;
Research Areas |signal transduction|protein phosphorylation|ser / thr kinases|other kinases|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for Serine/threonine-protein kinase PLK2

Polo-like kinase2, also known as SNK is an enzyme that in humans is encoded by the PLK2 gene. PLK2 is a member of the "polo" family of serine/threonine protein kinases that have a role in normal cell division. The International Radiation Hybrid Mapping Consortium mapped the SNK gene to chromosome 5. SNK involved in synaptic plasticity, centriole duplication and G1/S phase transition. This gene plays a key role in synaptic plasticity and memory by regulating the Ras and Rap protein signalin: required for overactivity-dependent spine remodeling by phosphorylating the Ras activator RASGRF1 and the Rap inhibitor SIPA1L1 leading to their degradation by the proteasome.

Anti-PLK2/Snk Antibody Images

Click the images to enlarge.

Anti-PLK2/Snk Antibody
Figure 1. Western blot analysis of PLK2/Snk using anti- PLK2/Snk antibody (PA2131).
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 50ug of sample under reducing conditions.
Lane 1: A431 Cell Lysate
Lane 2: 293T Cell Lysate
Lane 3: COLO-320 Cell Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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- PLK2/Snk antigen affinity purified polyclonal antibody (Catalog # PA2131) 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 1:10000 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 PLK2/Snk at approximately 78KD. The expected band size for PLK2/Snk is at 77KD.
Anti-PLK2/Snk Antibody
Figure 2. IHC analysis of PLK2/Snk using anti- PLK2/Snk antibody (PA2131).
PLK2/Snk was detected in paraffin-embedded section of human lung cancer tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- PLK2/Snk Antibody (PA2131) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-PLK2/Snk Antibody
Figure 3. IHC analysis of PLK2/Snk using anti- PLK2/Snk antibody (PA2131).
PLK2/Snk was detected in paraffin-embedded section of rat intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- PLK2/Snk Antibody (PA2131) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-PLK2/Snk Antibody
Figure 4. IHC analysis of PLK2/Snk using anti- PLK2/Snk antibody (PA2131).
PLK2/Snk was detected in paraffin-embedded section of rat brain tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti- PLK2/Snk Antibody (PA2131) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-PLK2/Snk Antibody
Figure 5. Flow Cytometry analysis of A549 cells using anti-PLK2 antibody (PA2131).
Overlay histogram showing A549 cells stained with PA2131 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-PLK2 Antibody (PA2131,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Customer Q&As

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Is your western blot protocol provided from the website applicable for all your antibodies? Keyword: applications, WB
A: The protocol is applicable for all our antibodies in WB, the NC Membrane(0.45μm or 0.22μm) and transfer time(70 mins or 50 mins) depends on the protein molecular weight, details can be found in included protocol.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands

3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.

4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.