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Product Info Summary
| SKU: | A01390-2 |
|---|---|
| Size: | 100 µg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, WB |
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Product info
Product Name
Anti-SPHK1 Antibody Picoband®
View all Sphingosine Kinase 1/SPHK1 Antibodies
SKU/Catalog Number
A01390-2
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-SPHK1 Antibody Picoband® catalog # A01390-2. Tested in ELISA, Flow Cytometry, WB applications. This antibody reacts with Human. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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-SPHK1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01390-2)
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 SPHK1 recombinant protein (Position: E55-N357).
*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
A01390-2 is reactive to SPHK1 in Human
Reconstitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Observed Molecular Weight
52 kDa
Calculated molecular weight
102380 MW
Background of Sphingosine Kinase 1/SPHK1
SPHK1(Sphingosine Kinase 1), is an enzyme that in humans is encoded by the SPHK1 gene. Melendez et al.(2000) mapped the SPHK1 gene to chromosome 17q25.2 based on sequence identity with ESTs mapped to this region. Kohama et al.(1998) demonstrated that recombinant mouse Sphk1 can specifically phosphorylate D-erythro-sphingosine and that D, L-threo-dihydrosphingosine and N, N-dimethylsphingosine can act as competitive inhibitors of recombinant Sphk1. Pitson et al.(2000) found that recombinant SPHK1 and endogenous SPHK1 purified from placenta had identical enzymatic characteristics, suggesting posttranslational modification does not effect functional properties.
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.
Application & Images
Applications
A01390-2 is guaranteed for ELISA, Flow Cytometry, WB Boster Guarantee
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
Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: human Hela whole cell, human HepG2 whole cell, human MCF-7 whole cell
FCM: HepG2 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of SPHK1 using anti-SPHK1 antibody (A01390-2).
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 Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human MCF-7 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-SPHK1 antigen affinity purified polyclonal antibody (Catalog # A01390-2) 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 SPHK1 at approximately 52 kDa. The expected band size for SPHK1 is at 42 kDa.
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Figure 2. Flow Cytometry analysis of HepG2 cells using anti-SPHK1 antibody (A01390-2).
Overlay histogram showing HepG2 cells stained with A01390-2 (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-SPHK1 Antibody (A01390-2, 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Protein Target Info & Infographic
Gene/Protein Information For SPHK1 (Source: Uniprot.org, NCBI)
Gene Name
SPHK1
Full Name
Sphingosine kinase 1
Weight
102380 MW
Alternative Names
Hexokinase-2;2.7.1.1;Hexokinase type II;HK II;Muscle form hexokinase;HK2; SPHK1 SPHK sphingosine kinase 1 sphingosine kinase 1|SK 1|SPK 1|acetyltransferase SPHK1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on SPHK1, check out the SPHK1 Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for SPHK1: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-SPHK1 Antibody Picoband® (A01390-2)
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