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Product Info Summary
| SKU: | M02794 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IHC, IHC-F, ICC, WB |
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Product info
Product Name
Anti-Stefin B CSTB Antibody Picoband® (monoclonal, 2B6)
SKU/Catalog Number
M02794
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Stefin B CSTB Antibody Picoband® (monoclonal, 2B6) catalog # M02794. Tested in Flow Cytometry, IHC, IHC-F, ICC, 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
Store 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 freeze-thaw cycles.
Cite This Product
Anti-Stefin B CSTB Antibody Picoband® (monoclonal, 2B6) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M02794)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
2B6
Isotype
Mouse IgG1
Immunogen
E. coli-derived human Stefin B recombinant protein (Position: M1-F98). Human Stefin B shares 78.6 % amino acid (aa) sequence identity with both mouse and rat Stefin B.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M02794 is reactive to CSTB in Human
Observed Molecular Weight
14 kDa
Calculated molecular weight
11.1 kDa
Background of CSTB
Cystatin B (CSTB), also called STFB, is a small protein that is a member of the superfamily of cysteine protease inhibitors. It has been isolated from human spleen and liver and its amino acid sequence has been fully determined. The cystatin B gene is located on 21q22.3. It is widely distributed and is localized mostly intracellularly, but has been found extracellularly. The protein is able to form a dimer stabilized by noncovalent forces, inhibiting papain and cathepsins l, h and b. Its role is thought to be as a protector against the proteinases leaking from lysosomes. A cystatin B multiprotein complex might have a specific cerebellar function, and that the loss of this function might contribute to the etiopathogenesis of EPM1. Upon differentiation to myotubes, CSTB becomes excluded from the nucleus and lysosomes, suggesting that the subcellular distribution of CSTB is dependent on the differentiation status of the cell.
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
M02794 is guaranteed for Flow Cytometry, IHC, IHC-F, ICC, 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, 0.1-0.5μg/ml
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Immunohistochemistry (Frozen Section), 0.5-1μg/ml
Immunocytochemistry, 0.5-1μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
Positive Control
WB: human U-87MG whole cell, human SW620 whole cell, human U-937 whole cell, human Caco-2 whole cell
IHC: human tonsil tissue
FCM: PC-3 cell
Validation Images & Assay Conditions
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Western blot analysis of Stefin B using anti-Stefin B antibody (M02794).
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 U-87MG whole cell lysates,
Lane 2: human SW620 whole cell lysates,
Lane 3: human U-937 whole cell lysates,
Lane 4: human Caco-2 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 mouse anti-Stefin B antigen affinity purified monoclonal antibody (Catalog # M02794) 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-mouse 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 # EK1001) with Tanon 5200 system.
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IHC analysis of Stefin B using anti-Stefin B antibody (M02794).
Stefin B was detected in paraffin-embedded section of human tonsil tissue. 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 2μg/ml mouse anti-Stefin B Antibody (M02794) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.
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Flow Cytometry analysis of PC-3 cells using anti-Stefin B antibody (M02794).
Overlay histogram showing PC-3 cells stained with M02794 (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 mouse anti-Stefin B Antibody (M02794,1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Specific Publications For Anti-Stefin B CSTB Antibody Picoband® (monoclonal, 2B6) (M02794)
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