Product Info Summary
SKU: | M00676-5 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human |
Host: | Mouse |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Hsp27/HSPB1 Antibody Picoband™ (monoclonal, 3H3)
SKU/Catalog Number
M00676-5
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Hsp27/HSPB1 Antibody Picoband™ (monoclonal, 3H3) catalog # M00676-5. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human.
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-Hsp27/HSPB1 Antibody Picoband™ (monoclonal, 3H3) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00676-5)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
3H3
Isotype
Mouse IgG1
Immunogen
E.coli-derived human Hsp27/HSPB1 recombinant protein (Position: M1-K205). Human Hsp27 shares 83% amino acid (aa) sequence identity with mouse Hsp27.
*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
M00676-5 is reactive to HSPB1 in Human
Applications
M00676-5 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
27 kDa
Calculated molecular weight
22.783kDa
Background of HSP27
HSPB1 (Heat shock 27kDa protein 1), also known as HSP27, is a protein that in humans is encoded by the HSPB1 gene. HSP27 gene is mapped to 7q11.23. The protein encoded by this gene is induced by environmental stress and developmental changes. The encoded protein is involved in stress resistance and actin organization and translocates from the cytoplasm to the nucleus upon stress induction. Defects in this gene are a cause of Charcot-Marie-Tooth disease type 2F (CMT2F) and distal hereditary motor neuropathy (dHMN).
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, 2μg/ml
Flow Cytometry, 1-3μg/1x106 cells
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of HSP27 using anti-HSP27 antibody (M00676-5).
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 CACO-2 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 mouse anti-HSP27 antigen affinity purified monoclonal antibody (Catalog # M00676-5) 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 HSP27 at approximately 27 kDa. The expected band size for HSP27 is at 27 kDa.
Click image to see more details
Figure 2. IHC analysis of HSP27 using anti-HSP27 antibody (M00676-5).
HSP27 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-HSP27 Antibody (M00676-5) 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 HSP27 using anti-HSP27 antibody (M00676-5).
HSP27 was detected in paraffin-embedded section of human oesophagus squama 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-HSP27 Antibody (M00676-5) 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 HSP27 using anti-HSP27 antibody (M00676-5).
HSP27 was detected in paraffin-embedded section of human placenta 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-HSP27 Antibody (M00676-5) 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. IF analysis of HSP27 using anti- HSP27 antibody (M00676-5).
HSP27 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 mouse anti- HSP27 Antibody (M00676-5) 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.
Click image to see more details
Figure 6. Flow Cytometry analysis of A549 cells using anti-HSP27 antibody (M00676-5).
Overlay histogram showing A549 cells stained with M00676-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSP27 Antibody (M00676-5,1μg/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 7. Flow Cytometry analysis of U20S cells using anti-HSP27 antibody (M00676-5).
Overlay histogram showing U20S cells stained with M00676-5 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSP27 Antibody (M00676-5,1μg/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.
Protein Target Info & Infographic
Gene/Protein Information For HSPB1 (Source: Uniprot.org, NCBI)
Gene Name
HSPB1
Full Name
Heat shock protein beta-1
Weight
22.783kDa
Superfamily
small heat shock protein (HSP20) family
Alternative Names
28 kDa heat shock protein; DKFZp586P1322; Estrogen-regulated 24 kDa protein; Heat shock 27 kDa protein; heat shock 27kD protein 1; heat shock 27kDa protein 1; heat shock protein beta-1; HMN2B; HS.76067; HSP25; HSP27; HSP27HSP 27; HSP28CMT2F; HSPB1; SRP27; Stress-responsive protein 27 HSPB1 CMT2F, HEL-S-102, HMN2B, HS.76067, HSP27, HSP28, Hsp25, SRP27 heat shock protein family B (small) member 1 heat shock protein beta-1|28 kDa heat shock protein|epididymis secretory protein Li 102|estrogen-regulated 24 kDa protein|heat shock 27 kDa protein|heat shock 27kD protein 1|heat shock 27kDa protein 1|stress-responsive protein 27
*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 HSPB1, check out the HSPB1 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 HSPB1: 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-Hsp27/HSPB1 Antibody Picoband™ (monoclonal, 3H3) (M00676-5)
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