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Product Info Summary
| SKU: | M00961-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, IHC, ICC, WB, ELISA (Cap) |
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Product info
Product Name
Anti-Cystatin C/CST3 Antibody Picoband® (monoclonal, 4H8)
SKU/Catalog Number
M00961-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Cystatin C/CST3 Antibody Picoband® (monoclonal, 4H8) catalog # M00961-1. Tested in ELISA, Flow Cytometry, IF, IHC, 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-Cystatin C/CST3 Antibody Picoband® (monoclonal, 4H8) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00961-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
4H8
Isotype
Mouse IgG2a
Immunogen
E. coli-derived human Cystatin C recombinant protein (Position: K31-A146).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00961-1 is reactive to CST3 in Human
Observed Molecular Weight
15 kDa
Calculated molecular weight
15.8 kDa
Background of CST3
Cystatin C or cystatin 3, a protein encoded by the CST3 gene, is mainly used as a biomarker of kidney function. Recently, it has been studied for its role in predicting new-onset or deteriorating cardiovascular disease. It also seems to play a role in brain disorders involving amyloid, such as Alzheimer's disease. In humans, all cells with a nucleus (cell core containing the DNA) produce cystatin C as a chain of 120 amino acids. It is found in virtually all tissues and body fluids. It is a potent inhibitor of lysosomal proteinases (enzymes from a special subunit of the cell that break down proteins) and probably one of the most important extracellular inhibitors of cysteine proteases (it prevents the breakdown of proteins outside the cell by a specific type of protein degrading enzymes). Cystatin C belongs to the type 2 cystatin gene family.
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
M00961-1 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB, ELISA (Cap) 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
Immunocytochemistry/Immunofluorescence, 2μg/ml
Flow Cytometry (Fixed), 1-3μg/1x106 cells
ELISA (Cap), 1-5μg/ml
Positive Control
WB: human PC-3 whole cell, human CACO-2 whole cell, human placenta tissue, human A549 whole cell, human COLO320 whole cell, human HEK293 whole cell
IHC: human colon cancer tissue, human colon cancer tissue, human glioma tissue
ICC/IF: MCF7 cell
FCM: A549 cell
Validation Images & Assay Conditions
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IF analysis of CST3 using anti-CST3 antibody (M00961-1).
CST3 was detected in immunocytochemical section of MCF7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-CST3 Antibody (M00961-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Western blot analysis of Cystatin C/CST3 using anti-Cystatin C/CST3 antibody (M00961-1).
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 PC-3 whole cell lysates,
Lane 2: human CACO-2 whole cell lysates,
Lane 3: human placenta tissue lysates,
Lane 4: human A549 whole cell lysates,
Lane 5: human COLO320 whole cell lysates,
Lane 6: human HEK293 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-Cystatin C/CST3 antigen affinity purified monoclonal antibody (Catalog # M00961-1) 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. A specific band was detected for Cystatin C/CST3 at approximately 15KD. The expected band size for Cystatin C/CST3 is at 15KD.
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IHC analysis of Cystatin C/CST3 using anti-Cystatin C/CST3 antibody (M00961-1).
Cystatin C/CST3 was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cystatin C/CST3 Antibody (M00961-1) 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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IHC analysis of Cystatin C/CST3 using anti-Cystatin C/CST3 antibody (M00961-1).
Cystatin C/CST3 was detected in paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cystatin C/CST3 Antibody (M00961-1) 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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IHC analysis of Cystatin C/CST3 using anti-Cystatin C/CST3 antibody (M00961-1).
Cystatin C/CST3 was detected in paraffin-embedded section of human glioma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-Cystatin C/CST3 Antibody (M00961-1) 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 A549 cells using anti-Cystatin C/CST3 antibody (M00961-1).
Overlay histogram showing A549 cells stained with M00961-1 (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-Cystatin C/CST3 Antibody (M00961-1, 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.
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Sandwich ELISA - Recombinant human Cystatin C/CST3 protein standard curve.
Use in combination with reagents from Human Cystatin C/CST3 ELISA Kit EZ-Set (DIY Antibody Pairs) (EZ0678).
Specific Publications For Anti-Cystatin C/CST3 Antibody Picoband® (monoclonal, 4H8) (M00961-1)
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