Anti-phospho-HSP27 Ser82 Antibody, clone 1O17 ZooMAb® Rabbit Monoclonal
SIGMA/ZRB1659 - recombinant, expressed in HEK 293 cells
Synonym: Heat shock protein beta-1
Product Type: Chemical
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in muscularis mucosae of Human Small Intestine (image set 2). Membranous staining was also observed in Human Colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in muscularis mucosae of Human Small Intestine (image set 2). Membranous staining was also observed in Human Colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Colon cancer (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Membranous staining was observed in muscularis mucosae of Human Small Intestine (image set 2). Membranous staining was also observed in Human Colon cancer tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Small Intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of glandular cells, lamina propria & blood vessels of submucosa of Human Small Intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Small Intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of glandular cells, lamina propria & blood vessels of submucosa of Human Small Intestine tissue sections.
Immunohistochemistry (Paraffin) Enhanced Validation - Recombinant Antibody Technology Formalin Fixed Paraffin Embedded (FFPE) Human Small Intestine (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in subsets of glandular cells, lamina propria & blood vessels of submucosa of Human Small Intestine tissue sections.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with Anisomycin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus, Cytoplasm, and Cytoskeleton.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with Anisomycin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus, Cytoplasm, and Cytoskeleton.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with Anisomycin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus, Cytoplasm, and Cytoskeleton.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with Anisomycin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus, Cytoplasm, and Cytoskeleton.
Immunocytochemistry Enhanced Validation - Recombinant Antibody Technology Immunofluorescent analysis of untreated HeLa cells (A and B) and HeLa cells treated with Anisomycin (C and D) were performed using a 1:100 dilution of Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal and visualized with a Goat Anti-Rabbit secondary antibody conjugated to Alexa Fluor® 488 (Green). Actin filaments have been labeled with phalloidin (Red). Nucleus is stained with DAPI (Blue). This antibody positively stains the Nucleus, Cytoplasm, and Cytoskeleton.
Western Blotting Enhanced Validation - Recombinant Antibody Technology Lysates from untreated HeLa cells (Lane 1) and UV-treated HeLa cells (Lane 2) were probed with Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-HSP27-Ser82 (~30 kDa).
Affinity Binding Assay Enhanced Validation - Recombinant Antibody Technology 100 mg/mL of non-phospho HSP27 control peptide (Run 1) or phospho-HSP27-Ser82 peptide (Run 2) were analyzed by affinity binding assay using Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal. The binding of this antibody to phospho-HSP27-Ser82 peptide displayed a KD of 1 x 10-7, whereas binding to non-phospho HSP27 control peptide displayed a KD of 5.9 x 10-4.
Peptide Inhibition Assay Enhanced Validation - Recombinant Antibody Technology Lysates from HeLa treated with Anisomycin was probed with Cat. No. ZRB1659, Anti-phospho-HSP27 Ser82, clone 1O17 ZooMAb® Rabbit Monoclonal (1:10,000 dilution). Proteins were visualized using a Donkey Anti-Rabbit IgG secondary antibody conjugated to HRP and a chemiluminescence detection system. Arrow indicates phospho-HSP27-Ser82 (~30 kDa). Lanes 1: Incubated without any peptide. 2: Incubated with phospho-HSP27 Ser82 peptide. 3: Incubated with unphosphorylated HSP-27 peptide.
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Chemical products should be designed to affect their desired function while minimizing their toxicity.
12 Principles of Green Chemistry: Principle 6—Design for Energy Efficiency. This product was re-engineered to utilize less energy in the manufacturing process.
| antibody form | purified antibody |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | 1O17, recombinant monoclonal |
| conjugate | unconjugated |
| description | recombinant, expressed in HEK 293 cells |
| enhanced validation | recombinant expression Learn more about Antibody Enhanced Validation  |
| epitope sequence | N-terminal half |
| form | lyophilized |
| greener alternative category | Aligned |
| greener alternative product characteristics | Waste Prevention Designing Safer Chemicals Design for Energy Efficiency Learn more about the Principles of Green Chemistry . |
| isotype | IgG |
| mol wt | calculated mol wt 22.78 kDa |
| observed mol wt ~30 kDa | |
| packaging | antibody small pack of 25 μL |
| product line | ZooMAb® learn more |
| Protein ID accession no. | NP_001531  |
| purified by | using Protein A |
| Quality Level | 200 |
| recombinant | expressed in HEK 293 cells |
| shipped in | ambient |
| species reactivity | human |
| species reactivity (predicted by homology) | bovine, horse, mouse, feline, monkey, canine, rat |
| storage temp. | 2-8°C |
| UniProt accession no. | P04792 |
| Application: | Quality Control Testing Evaluated by Western Blotting in lysates from HeLa cells overnight serum starved and treated with Anisomycin. Western Blotting Analysis (WB): A 1:10,000 dilution of this antibody detected HSP27 phosphorylated on serine 82 in lysates of HeLa cells overnight serum starved and treated with Anisomycin (25 μg/mL; 30 min.). Tested applications Western Blotting Analysis: A 1:10,000 dilution from a representative lot detected phospho-HSP27 Ser82 in lysate from UV-treated HeLa cells. Immunohistochemistry (Paraffin) Analysis: A 1:100 dilution from a representative lot detected phospho-HSP27 Ser82 in human small intestine and human colon cancer tissue sections. Affinity Binding Assay: A representative lot of this antibody bound phospho-HSP27-ser82 peptide with a KD of 1. x 10-7 in an affinity binding assay. Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho-HSP27 Ser82 in HeLa cells treated with Anisomycin. Peptide Inhibition Assay: Target band detection in lysate from HeLa cells treated with Anisomycin was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide. Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| General description: | We are committed to bringing you greener alternative products, which adhere to one or more of The 12 Principles of Green Chemistry.This antibody is Preservative-free, produced without the harm or sacrifice of animals and exceptionally stable to allow for ambient shipping and storage if needed and thus aligns with "Waste Prevention", "Designing Safer Chemicals" and "Design for Energy Efficiency". Click here for more information. |
| General description: | ZooMAb® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb® antibodies are reliably available and ready to ship when you need them. |
| Immunogen: | KLH-conjugated linear peptide corresponding to 11 amino acids surrounding phosphoserine 82 from the N-terminal half of human Heat shock protein 27 (HSP27). |
| Legal Information: | ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany |
| Physical form: | Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 μL. |
| Reconstitution: | 300 μg/mL after reconstitution at 25 μL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type. |
| Specificity: | Clone 1O17 is a ZooMAb® Rabbit recombinant monoclonal antibody that specifically detects HSP27 phosphorylated on serine 82. |
| Storage and Stability: | Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 μL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws. |
| Target description: | Heat shock protein beta-1 (UniProt: P04792; also known as HspB1, 28 kDa heat shock protein, Estrogen-regulated 24 kDa protein, Heat shock 27 kDa protein, HSP 27, Stress-responsive protein 27, SRP27) is encoded by the HSPB1 (also known as HSP27, HSP28) gene (Gene ID: 3315) in human. Heat shock proteins (HSPs) are produced during nonlethal stress conditions and protect the organism from severe and sometimes lethal stress. HSPs can also be induced by non-heat related stress factors. HSP27 is a multidimensional protein that acts as a protein chaperone and an antioxidant and plays a role in the inhibition of apoptosis and actin cytoskeletal remodeling. It is detected in all tissues; however, its highest levels are found in the heart and in tissues composed of striated and smooth muscle. It is shown to be mostly cytoplasmic in interphase cells and is shown to colocalize with mitotic spindles in mitotic cells. It translocates to the nucleus during heat shock and resides in sub-nuclear structures known as nuclear splicing speckles and functions to stabilize DNA and/or the nuclear membrane. HSP-27 contains a conserved -crystalline homology domain at its C-terminal. It is phosphorylated upon exposure to protein kinase C activators and heat shock. HSP27 is present at basal levels in cells and tissues and is organized as large oligomers. After heat shock, exposure to chemical stress, or addition of steroid hormones, its synthesis increases several fold. It is phosphorylated by MAPKAP kinase 2/3 via the activation of the p38 MAPK pathway at multiple serine residues. Following phosphorylation, HSP27 reorganizes itself into smaller oligomers that interact with other proteins. This ZooMAb® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, affinity, reproducibility, and stability over conventional monoclonals. (Ref.: Alderson TR., et al. (2019). Nat. Commun. 10; Article 1068; Vidyasagar, A., et al. (2012). Fibrogenesis Tissue Repair. 5(1); 7; Landry, J., et al. (1992). J. Biol. Chem. 267(2); 794-803). |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352203 |