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Product Info Summary
| SKU: | PB9337 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Rabbit |
| Application: | IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Hsp60/HSPD1 Antibody Picoband®
SKU/Catalog Number
PB9337
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hsp60/HSPD1 Antibody Picoband® catalog # PB9337. Tested in IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. 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-Hsp60/HSPD1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9337)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human Hsp60 recombinant protein (Position: A260-Q496). Human Hsp60 shares 97% amino acid (aa) sequence identity with both mouse and rat Hsp60.
Cross-reactivity
No cross-reactivity with other proteins
Reactive Species
PB9337 is reactive to HSPD1 in Human, Mouse, Rat
Observed Molecular Weight
61 kDa
Calculated molecular weight
61.1 kDa
Background of HSPD1
HSP60 is a member of the chaperonin class of protein factors, which include the Escherichia coli groEL protein and the Rubisco subunit-binding protein of chloroplasts. It acts as a costimulator of human regulatory CD4-positive/CD25 -positive T cells, which inhibit lymphoproliferation and IFNG and TNF secretion by CD4-positive and CD8-positive T cells. HSP60 enhances Treg activity via TLR2, leading to activation of an intracellular signaling cascade that included p38, as well as inhibition of ERK phosphorylation. Suppression of target T cells is mediated by both cell-to-cell contact and by secretion of TGFB and IL10, and it leads to downregulation of ERK, NFKB, and TBET expression. The self-molecule HSP60 can downregulate adaptive immune responses by upregulating Tregs through TLR2 signaling.
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
PB9337 is guaranteed for IF, IHC, 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, Human, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Positive Control
WB: human Hela whole cell, human A549 whole cell, human PANC-1 whole cell, human MCF-7 whole cell, rat liver tissue, mouse liver tissue, mouse thymus tissue
IHC: Mouse Intestine Tissue, Rat Intestine Tissue, Human Intestinal Cancer Tissue
ICC/IF: U20S cell
ICC: SMMC-7721 cell
Validation Images & Assay Conditions
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Western blot analysis of Hsp60 using anti-Hsp60 antibody (PB9337).
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 A549 whole cell lysates,
Lane 3: human PANC-1 whole cell lysates,
Lane 4: human MCF-7 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: mouse liver tissue lysates,
Lane 7: mouse thymus tissue 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-Hsp60 antigen affinity purified polyclonal antibody (Catalog # PB9337) 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 Hsp60 at approximately 61 kDa. The expected band size for Hsp60 is at 61 kDa.
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Anti-Hsp60 Picoband antibody, PB9337, IHC(P)
IHC(P): Human Intestinal Cancer Tissue
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Anti-Hsp60 Picoband antibody, PB9337, IHC(P)
IHC(P): Rat Intestine Tissue
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Anti-Hsp60 Picoband antibody, PB9337, IHC(P)
IHC(P): Mouse Intestine Tissue
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IHC analysis of Hsp60 using anti-Hsp60 antibody (PB9337).
Hsp60 was detected in immunocytochemical section of SMMC-7721 Cell. 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 1μg/ml rabbit anti-Hsp60 Antibody (PB9337) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
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IF analysis of Hsp60 using anti-Hsp60 antibody (PB9337).
Hsp60 was detected in immunocytochemical section of U20S cell. 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 rabbit anti-Hsp60 Antibody (PB9337) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.
Specific Publications For Anti-Hsp60/HSPD1 Antibody Picoband® (PB9337)
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Customer Q&As
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2 Customer Q&As for Anti-Hsp60/HSPD1 Antibody Picoband®
Question
Is PB9337 suitable for IHC frozen section staining?
Verified customer
Asked: 2019-04-23
Answer
The Anti-Hsp60/HSPD1 Antibody Picoband™ (PB9337) has been validated for IHC-F on human samples. The Suggested dilution for Immunohistochemistry (Frozen Section) is 0.5-1μg/ml, Human
Boster Scientific Support
Answered: 2019-04-23
Question
We are currently using anti-Hsp60/HSPD1 antibody PB9337 for human tissue, and we are happy with the IHC-P results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on dog tissues as well?
B. Singh
Verified customer
Asked: 2015-12-08
Answer
The anti-Hsp60/HSPD1 antibody (PB9337) has not been validated for cross reactivity specifically with dog tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2015-12-08
