Product Info Summary
SKU: | M01280-3 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Mouse |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Hsp60/HSPD1 Antibody Picoband™ (monoclonal, 6G2)
SKU/Catalog Number
M01280-3
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hsp60/HSPD1 Antibody Picoband™ (monoclonal, 6G2) catalog # M01280-3. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
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™ (monoclonal, 6G2) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M01280-3)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
6G2
Isotype
Mouse IgG1
Immunogen
E.coli-derived human Hsp60/HSPD1 recombinant protein (Position: A260-Q496). Human Hsp60 shares 97% amino acid (aa) sequence identity with both mouse and rat Hsp60.
*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
M01280-3 is reactive to HSPD1 in Human, Mouse, Rat
Applications
M01280-3 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
60 kDa
Calculated molecular weight
61.055kDa
Background of HSP60
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.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
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, 5μg/ml
Flow Cytometry, 1-3μg/1x106 cells
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
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 Caco-2 whole cell lysates
Lane 2: human A549 whole cell lysates
Lane 3: human THP-1 whole cell lysates
Lane 4: human SW620 whole cell lysates
Lane 5: human U-937 whole cell lysates
Lane 6: human HepG2 whole cell lysates
Lane 7: rat RH35 whole cell lysates
Lane 8: mouse RAW246.7 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-HSPD1 antigen affinity purified monoclonal antibody (Catalog # M01280-3) 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 HSPD1 at approximately 60KD. The expected band size for HSPD1 is at 60KD.
Click image to see more details
Figure 2. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of human intestinal cancer tissues. 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 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) 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.
Click image to see more details
Figure 3. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of human lung cancer tissues. 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 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) 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.
Click image to see more details
Figure 4. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of human mammary cancer tissues. 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 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) 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.
Click image to see more details
Figure 5. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of mouse liver tissues. 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 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) 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.
Click image to see more details
Figure 6. IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of rat liver tissues. 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 1μg/ml mouse anti-HSPD1 Antibody (M01280-3) 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.
Click image to see more details
Figure 7. Flow Cytometry analysis of A431 cells using anti-HSPD1 antibody (M01280-3).
Overlay histogram showing A431 cells stained with M01280-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSPD1 Antibody (M01280-3,1μg/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.
Click image to see more details
Figure 8. Flow Cytometry analysis of HepG2 cells using anti-HSPD1 antibody (M01280-3).
Overlay histogram showing HepG2 cells stained with M01280-3 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSPD1 Antibody (M01280-3,1μg/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.
Click image to see more details
Figure 9. IF analysis of Hsp60/HSPD1 using anti-Hsp60/HSPD1 antibody (M01280-3).
Hsp60/HSPD1 was detected in immunocytochemical section of A431 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 5μg/mL mouse anti-Hsp60/HSPD1 Antibody (M01280-3) 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.
Protein Target Info & Infographic
Gene/Protein Information For HSPD1 (Source: Uniprot.org, NCBI)
Gene Name
HSPD1
Full Name
60 kDa heat shock protein, mitochondrial
Weight
61.055kDa
Superfamily
chaperonin (HSP60) family
Alternative Names
60 kDa chaperonin; Chaperonin 60; cpn60; GROEL; heat shock 60kD protein 1 (chaperonin); heat shock 60kDa protein 1 (chaperonin); Heat shock protein 60; heat shock protein 65; HLD4; HSP60; HSP-60; HSP60SPG13; HSP65; HSPD1; HuCHA60; Mitochondrial matrix protein P1,60 kDa heat shock protein, mitochondrial; P60 lymphocyte protein; short heat shock protein 60 Hsp60s1; spastic paraplegia 13 (autosomal dominant); SPG13 HSPD1 CPN60, GROEL, HLD4, HSP-60, HSP60, HSP65, HuCHA60, SPG13 heat shock protein family D (Hsp60) member 1 60 kDa heat shock protein, mitochondrial|60 kDa chaperonin|P60 lymphocyte protein|chaperonin 60|epididymis secretory sperm binding protein|heat shock 60kDa protein 1 (chaperonin)|heat shock protein 65|mitochondrial matrix protein P1|short heat shock protein 60 Hsp60s1
*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 HSPD1, check out the HSPD1 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 HSPD1: 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-Hsp60/HSPD1 Antibody Picoband™ (monoclonal, 6G2) (M01280-3)
Hello CJ!
M01280-3 has been cited in 3 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Expression and location of HSP60 and HSP10 in the heart tissue of heat-stressed rats
Hepatic mitochondrial and ER stress induced by defective PPAR? signaling in the pathogenesis of hepatic steatosis
Hager L, Li L, Pun H, Liu L, Hossain Ma, Maguire Gf, Naples M, Baker C, Magomedova L, Tam J, Adeli K, Cummins Cl, Connelly Pw, Ng Ds. J Biol Chem. 2012 Jun 8;287(24):20755-68. Doi: 10.1074/Jbc.M112.340919. Epub 2012 Apr 12. Lecithin:Cholesterol Ac...
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