Recombinant Anti-SPAK antibody [EPR6394] (ab128894)
![Western blot - Anti-SPAK antibody [EPR6394] (ab128894)](https://www.abcam.com/ps/products/128/ab128894/Images/ab128894-275416-anti-stk39-antibody-epr6394-western-blot.jpg "Western blot - Anti-SPAK antibody [EPR6394] (ab128894)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6394] to SPAK
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-SPAK antibody [EPR6394]
See all SPAK primary antibodies -
Description
Rabbit monoclonal [EPR6394] to SPAK -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt (Intra),ICC/IF or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human SPAK aa 400-500.
Database link: Q9UEW8 -
Positive control
- SH-SY5Y, 293T, Jurkat, BxPC3 and HepG2 whole cell lysate (ab7900); Human colon tissue
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 3.10 x 10 -11 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR6394 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab128894 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 59 kDa).
|
| IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
| Notes |
|---|
|
WB
1/1000 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 59 kDa). |
|
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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Function
May act as a mediator of stress-activated signals. -
Tissue specificity
Predominantly expressed in brain and pancreas followed by heart, lung, kidney, skeletal muscle, liver, placenta and testis. -
Sequence similarities
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. STE20 subfamily.
Contains 1 protein kinase domain. -
Domain
PAPA box (proline-alanine repeats) may target the kinase to a specific subcellular location by facilitating interaction with intracellular proteins such as actin or actin-like proteins. -
Cellular localization
Cytoplasm. Nucleus. Nucleus when caspase-cleaved. - Information by UniProt
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Database links
- Entrez Gene: 27347 Human
- Omim: 607648 Human
- SwissProt: Q9UEW8 Human
- Unigene: 276271 Human
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Alternative names
- DCHT antibody
- DKFZp686K05124 antibody
- OTTHUMP00000165175 antibody
see all
Images
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Western blot - Anti-SPAK antibody [EPR6394] (ab128894)
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: SPAK knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jurkat whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab128894 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab128894 was shown to specifically react with SPAK in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when SPAK knockout samples were uexamined. Wild-type and SPAK knockout samples were subjected to SDS-PAGE. Ab128894 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging. -
![Western blot - Anti-SPAK antibody [EPR6394] (ab128894)](https://www.abcam.com/ps/products/128/ab128894/Images/ab128894-142695-anti-stk39-antibody-epr6394-western-blot.jpg)
Western blot - Anti-SPAK antibody [EPR6394] (ab128894)All lanes : Anti-SPAK antibody [EPR6394] (ab128894) at 1/1000 dilution
Lane 1 : SH-SY5Y cell lysate
Lane 2 : 293T (Human embryonic kidney epithelial cell) cell lysate
Lane 3 : Jurkat cell lysate
Lane 4 : BxPC-3 cell lysate
Lane 5 : HepG2 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 59 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted? -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPAK antibody [EPR6394] (ab128894)](https://www.abcam.com/ps/products/128/ab128894/Images/ab128894-142694-anti-stk39-antibody-epr6394-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SPAK antibody [EPR6394] (ab128894)ab128894, at 1/50 dilution, staining SPAK in paraffin embedded Human colon tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![OI-RD Scanning - Anti-SPAK antibody [EPR6394] (ab128894)](https://www.abcam.com/ps/products/128/ab128894/Images/ab128894-142693-anti-stk39-antibody-epr6394.jpg)
OI-RD Scanning - Anti-SPAK antibody [EPR6394] (ab128894) -
![Anti-SPAK antibody [EPR6394] (ab128894)](https://www.abcam.com/ps/products/128/ab128894/Images/ab128894-5-benefits-of-recombinant-antibodies.png)
Anti-SPAK antibody [EPR6394] (ab128894)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (2)
ab128894 has been referenced in 2 publications.
- Li C et al. MicroRNA-299-5p inhibits cell metastasis in breast cancer by directly targeting serine/threonine kinase 39. Oncol Rep 43:1221-1233 (2020). PubMed: 32020227
- Wolley MJ et al. In Primary Aldosteronism, Mineralocorticoids Influence Exosomal Sodium-Chloride Cotransporter Abundance. J Am Soc Nephrol 28:56-63 (2017). Human . PubMed: 27381844