![Knockout Validated: HSP27 Antibody (6H11) [NBP2-25149] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout (KO) HeLa cell line. PVDF membrane was probed with 1:5,000 of Mouse Anti-Human HSP27 Monoclonal Antibody (Catalog # NBP2-25149) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (catalog number HAF018). Specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.](http://images.novusbio.com/fullsize/HSP27-Antibody-6H11-Knockout-Validated-NBP2-25149-img0006.jpg "Knockout Validated: HSP27 Antibody (6H11) [NBP2-25149] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout (KO) HeLa cell line. PVDF membrane was probed with 1:5,000 of Mouse Anti-Human HSP27 Monoclonal Antibody (Catalog # NBP2-25149) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (catalog number HAF018). Specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.")
| Reactivity | Hu, Mu(-), Rt(-)Species Glossary |
| Applications | WB, Flow, ICC/IF, IHC, CyTOF-ready, KO |
| Clone | 6H11 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Concentration | 1 mg/ml |
| Immunogen | Recombinant full length human HSP27 expressed in and purified from E. coli. [UniProt# P04792] |
| Localization | Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > spindle. |
| Isotype | IgG1 |
| Clonality | Monoclonal |
| Host | Mouse |
| Gene | HSPB1 |
| Purity | Immunogen affinity purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
| Dilutions |
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| Application Notes | This Hsp27 (6H11) antibody is useful for Immunocytochemistry/Immunofluorescence and Western Blot, where a band is seen at ~27 kDa. This antibody is CyTOF ready. |
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| Theoretical MW | 27 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | 50% PBS, 50% glycerol |
| Preservative | 0.035% Sodium Azide |
| Concentration | 1 mg/ml |
| Purity | Immunogen affinity purified |
Secondary Antibodies |
Isotype Controls |
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![Western Blot: HSP27 Antibody (6H11) [NBP2-25149] - Analysis of tissue or whole cell lysates using mouse mAb to HSP27, NBP2-25149, dilution 1:10,000, in green. [1] protein standard (red), [2] rat brain, [3] mouse brain, [4] NIH-3T3, [5] HEK293, [6] HeLa, [7] SH-SY5Y cells. The strong single band at ~27 kDa corresponds to the HSP27 protein, detected only in human cell lines, since this antibody is human specific and unreactive with rodent HSP27. The blot was simultaneously probed with rabbit pAb to HSP-60, dilution 1:5,000 in red. A strong 60 kDa band is present in all preparations.](http://images.novusbio.com/fullsize/HSP27-Antibody-6H11-Western-Blot-NBP2-25149-img0007.jpg "Western Blot: HSP27 Antibody (6H11) [NBP2-25149] - Analysis of tissue or whole cell lysates using mouse mAb to HSP27, NBP2-25149, dilution 1:10,000, in green. [1] protein standard (red), [2] rat brain, [3] mouse brain, [4] NIH-3T3, [5] HEK293, [6] HeLa, [7] SH-SY5Y cells. The strong single band at ~27 kDa corresponds to the HSP27 protein, detected only in human cell lines, since this antibody is human specific and unreactive with rodent HSP27. The blot was simultaneously probed with rabbit pAb to HSP-60, dilution 1:5,000 in red. A strong 60 kDa band is present in all preparations.")
![Immunocytochemistry/Immunofluorescence: HSP27 Antibody (6H11) [NBP2-25149] - Analysis of HeLa cells stained with mouse HSP27 mAb, dilution 1:2000 (Green), and costained with chicken HSP60 pAb, dilution 1:2000 (Red). DAPI staining of nuclear DNA (Blue). The HSP27 antibody produces strong cytoplasm staining, while the HSP60 antibody specifically labels mitochondria](http://images.novusbio.com/fullsize/HSP27-Antibody-6H11-Immunocytochemistry-Immunofluorescence-NBP2-25149-img0008.jpg "Immunocytochemistry/Immunofluorescence: HSP27 Antibody (6H11) [NBP2-25149] - Analysis of HeLa cells stained with mouse HSP27 mAb, dilution 1:2000 (Green), and costained with chicken HSP60 pAb, dilution 1:2000 (Red). DAPI staining of nuclear DNA (Blue). The HSP27 antibody produces strong cytoplasm staining, while the HSP60 antibody specifically labels mitochondria")
![Flow Cytometry: Hsp27 Antibody (6H11) [NBP2-25149] - Flow cytometric staining of 1 x 10^6 MCF-7 cells using Hsp27 antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.](http://images.novusbio.com/fullsize/Hsp27-Antibody-6H11-Flow-Cytometry-NBP2-25149-img0005.jpg "Flow Cytometry: Hsp27 Antibody (6H11) [NBP2-25149] - Flow cytometric staining of 1 x 10^6 MCF-7 cells using Hsp27 antibody (dark blue). Isotype control shown in orange. An antibody concentration of 1 ug/1x10^6 cells was used.")
![Immunocytochemistry/Immunofluorescence: Hsp27 Antibody (6H11) [NBP2-25149] - HeLa cells stained with NBP2-25149 (red), and counterstained with chicken polyclonal antibody to Vimentin NB300-223 (green) and DNA (blue). NBP2-25149 reveals strong cytoplasmic staining and penetrates into the Actin rich ruffled margins, while the Vimentin antibody reveals cytoplasmic intermediate filaments.](http://images.novusbio.com/fullsize/Hsp27-Antibody-6H11-Immunocytochemistry-Immunofluorescence-NBP2-25149-img0003.jpg "Immunocytochemistry/Immunofluorescence: Hsp27 Antibody (6H11) [NBP2-25149] - HeLa cells stained with NBP2-25149 (red), and counterstained with chicken polyclonal antibody to Vimentin NB300-223 (green) and DNA (blue). NBP2-25149 reveals strong cytoplasmic staining and penetrates into the Actin rich ruffled margins, while the Vimentin antibody reveals cytoplasmic intermediate filaments.")
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![SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.](https://images.novusbio.com/images/Mouse-IgG1-Isotype-Control-MG1-SDS-Page-NBP1-97005-img0002.jpg "SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.")