Recombinant HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
![Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)](https://www.abcam.com/ps/products/194/ab194079/Images/ab194079-312035-ab194079KOWBJRIZ.jpg "Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- HRP Rabbit monoclonal [EPR5477] to Hsp27
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
- Conjugation: HRP
Overview
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Product name
HRP Anti-Hsp27 antibody [EPR5477]
See all Hsp27 primary antibodies -
Description
HRP Rabbit monoclonal [EPR5477] to Hsp27 -
Host species
Rabbit -
Conjugation
HRP -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat
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Immunogen
Synthetic peptide within Human Hsp27 aa 100-200. The exact sequence is proprietary.
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Positive control
- WB: HeLa cell lysate. IHC: normal human skeletal muscle tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C. Avoid freeze / thaw cycle. Store In the Dark. -
Storage buffer
pH: 7.40
Preservative: 0.1% 10% Proclin 300 Solution
Constituents: 30% Glycerol, 1% BSA, PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5477 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
Our Abpromise guarantee covers the use of ab194079 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-P | 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. ab199507 - Rabbit monoclonal IgG (HRP), is suitable for use an as isotype control with this antibody. |
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| WB | 1/5000. Detects a band of approximately 27 kDa (predicted molecular weight: 23 kDa). |
Target
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Function
Involved in stress resistance and actin organization. -
Tissue specificity
Detected in all tissues tested: skeletal muscle, heart, aorta, large intestine, small intestine, stomach, esophagus, bladder, adrenal gland, thyroid, pancreas, testis, adipose tissue, kidney, liver, spleen, cerebral cortex, blood serum and cerebrospinal fluid. Highest levels are found in the heart and in tissues composed of striated and smooth muscle. -
Involvement in disease
Defects in HSPB1 are the cause of Charcot-Marie-Tooth disease type 2F (CMT2F) [MIM:606595]. CMT2F is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. Nerve conduction velocities are normal or slightly reduced. CMT2F onset is between 15 and 25 years with muscle weakness and atrophy usually beginning in feet and legs (peroneal distribution). Upper limb involvement occurs later. CMT2F inheritance is autosomal dominant.
Defects in HSPB1 are a cause of distal hereditary motor neuronopathy type 2B (HMN2B) [MIM:608634]. Distal hereditary motor neuronopathies constitute a heterogeneous group of neuromuscular disorders caused by selective impairment of motor neurons in the anterior horn of the spinal cord, without sensory deficit in the posterior horn. The overall clinical picture consists of a classical distal muscular atrophy syndrome in the legs without clinical sensory loss. The disease starts with weakness and wasting of distal muscles of the anterior tibial and peroneal compartments of the legs. Later on, weakness and atrophy may expand to the proximal muscles of the lower limbs and/or to the distal upper limbs. -
Sequence similarities
Belongs to the small heat shock protein (HSP20) family. -
Post-translational
modificationsPhosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock. -
Cellular localization
Cytoplasm. Nucleus. Cytoplasm > cytoskeleton > spindle. Cytoplasmic in interphase cells. Colocalizes with mitotic spindles in mitotic cells. Translocates to the nucleus during heat shock and resides in sub-nuclear structures known as SC35 speckles or nuclear splicing speckles. - Information by UniProt
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Database links
- Entrez Gene: 3315 Human
- Entrez Gene: 15507 Mouse
- Entrez Gene: 24471 Rat
- Omim: 602195 Human
- SwissProt: P04792 Human
- SwissProt: P14602 Mouse
- SwissProt: P42930 Rat
- Unigene: 520973 Human
see all -
Alternative names
- Heat shock 27kDa protein antibody
- 28 kDa heat shock protein antibody
- CMT2F antibody
see all
Images
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Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)All lanes : HRP Anti-Hsp27 antibody [EPR5477] (ab194079) at 1/5000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : HSPB1 (Hsp27) knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsab194079 was shown to specifically react with Hsp27 in wild-type HAP1 cells as signal was lost in HSPB1 (Hsp27) knockout cells. Wild-type and HSPB1 (Hsp27) knockout samples were subjected to SDS-PAGE. Ab194079 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)](https://www.abcam.com/ps/products/194/ab194079/Images/ab194079-237306-IHCAP21158781.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)IHC image of Hsp27 staining in a section of formalin-fixed paraffin-embedded normal human skeletal muscle tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab194079 at 1/500 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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![Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)](https://www.abcam.com/ps/products/194/ab194079/Images/ab194079-232706-WBAP21158781.jpg)
Western blot - HRP Anti-Hsp27 antibody [EPR5477] (ab194079)HRP Anti-Hsp27 antibody [EPR5477] (ab194079) at 1/5000 dilution +HeLa whole cell lysate (ab150035) at 10 µg
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?
Exposure time: 20 secondsThis blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab194079 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.
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![HRP Anti-Hsp27 antibody [EPR5477] (ab194079)](https://www.abcam.com/ps/products/194/ab194079/Images/ab194079-4-benefits-of-recombinant-antibodies.png)
HRP Anti-Hsp27 antibody [EPR5477] (ab194079)
Protocols
References (0)
ab194079 has not yet been referenced specifically in any publications.
