Anti-MEKK2/MAP3K2 Antibody

SKU PA1499
Size 100μg/vial
Reactivity Human, Mouse, Rat
Clonality Polyclonal
Host Rabbit
Ig Isotype N/A
Applications WB

Overview

Product Name Anti-MEKK2/MAP3K2 Antibody
SKU/Catalog Number PA1499
Storage & Handling 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.
Size 100μg/vial
Description Rabbit IgG polyclonal antibody for Mitogen-activated protein kinase kinase kinase 2(MAP3K2) detection. Tested with WB in Human;Mouse;Rat.
Cite This Product Anti-MEKK2/MAP3K2 Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PA1499)
Host Rabbit
Contents/Buffer Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg Thimerosal, 0.05mg NaN3.
Form Lyophilized
Immunogen A synthetic peptide corresponding to a sequence at the C-terminus of human MEKK2(606-619aa SADELLRHMFVHYH), different from the related rat and mouse sequences by one amino acid.
Reactivity Human, Mouse, Rat

Assay Details

Assay Dilutions Overview

Concentration: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Western blot, 0.1-0.5μg/ml, Human, Rat, Mouse

Boster's Secondary Antibodies And IHC, WB Kits

The following reagents are used to generate the images below.

Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot.

Images And Assay Conditions

Figure 1. Western blot analysis of MEKK2 using anti-MEKK2 antibody (PA1499).
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: human Caco-2 whole cell lysates
Lane 2: human K562 whole cell lysates
Lane 3: human THP-1 whole cell lysates
Lane 4: human Raji whole cell lysates
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-MEKK2 antigen affinity purified polyclonal antibody (Catalog # PA1499) 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 MEKK2 at approximately 78KD. The expected band size for MEKK2 is at 70KD.

Figure 2. Western blot analysis of MEKK2 using anti-MEKK2 antibody (PA1499).
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: rat kidney tissue lysates
Lane 2: rat brain tissue lysates
Lane 3: rat heart tissue lysates
Lane 4: rat C6 whole cell lysates
Lane 5: mouse kidney tissue lysates
Lane 6: mouse brain tissue lysates
Lane 7: mouse heart tissue lysates
Lane 8: mouse Neuro-2a whole cell lysates
Lane 9: mouse NIH/3T3 whole cell lysates
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-MEKK2 antigen affinity purified polyclonal antibody (Catalog # PA1499) 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 MEKK2 at approximately 78KD. The expected band size for MEKK2 is at 70KD.

Target Info

Protein Target Info (Source: Uniprot.org)

Uniprot Id Q9Y2U5
Gene Name MAP3K2
Protein Name Mitogen-activated protein kinase kinase kinase 2
Alternative Names Mitogen-activated protein kinase kinase kinase 2;2.7.11.25;MAPK/ERK kinase kinase 2;MEK kinase 2;MEKK 2;MAP3K2;MAPKKK2, MEKK2;
Subcellular Localization Cytoplasm . Nucleus . Upon EGF stimulation, translocates into the nucleus.
Molecular Weight 69741 MW

*if product is indicated to react with multiple species, protein info is based on the human gene.

Ontology

Protein Function Component of a protein kinase signal transduction cascade. Regulates the JNK and ERK5 pathways by phosphorylating and activating MAP2K5 and MAP2K7 (By similarity). Plays a role in caveolae kiss-and-run dynamics. .
Research Areas Human, Mouse, Rat

*You can search these to find other products in these research areas.
Background MAP3K2, mitogen-activated protein kinase kinase kinase 2, also known as MEKK2, belongs to the mitogen-activated protein kinase(MAPK) kinase kinase gene family, which is involved in regulating multiple MAPK signaling pathways. Endogenous MAP3K2 localized mainly to the cytosol of unstimulated HeLa cells and rat fibroblasts. The MAP3K2 gene is mapped to chromosome 2 by the International Radiation Hybrid Mapping Consortium. MAP3K2 is a major upstream activator of the JNK, MAPK cascade. MAP3K2 was identifies as 1 of 6 kinases that regulate different steps of the caveolar cycle. Silencing of MAP3K2 strongly reduced kiss-and-run dynamics, leading to an accumulation of caveolar structures at the cell surface without affecting caveolae coat assembly or clustering.

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Polyclonal antibody for MEKK2/MAP3K2 detection. Host: Rabbit.Size: 100μg/vial. Tested applications: WB. Reactive species: Human. MEKK2/MAP3K2 information: Molecular Weight: 69741 MW; Subcellular Localization: Cytoplasm . Nucleus . Upon EGF stimulation, translocates into the nucleus.
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In stock
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PA1499
Buy one primary antibody get one 0.5ml HRP or Biotin secondary antibody for free.
*Sample sizes are prepared on demand and will take extra lead time. (cannot be conjugated)
$240.00

Troubleshooting

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Publications

Li Y, Zhang Z, Zhou X, Li L, Liu Q, Wang Z, Bai X, Zhao Y, Shi H, Zhang X, Ye L. Cancer Lett. 2014 Dec 28;355(2):288-96. Doi: 10.1016/J.Canlet.2014.09.047. Epub 2014 Oct 7. The Oncoprotein Hbxip Enhances Migration Of Breast Cancer Cells Through In...

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 "
  • 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:<br>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<br>2. Post-translational cleavage: this can cause smaller bands and or multiple bands <br><br>3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody. <br><br>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.<br>5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher., <br>
  • 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.
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