Recombinant Anti-Hsp27 antibody [EP1724Y] (ab62339)
![Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)](https://www.abcam.com/ps/products/62/ab62339/Images/ab62339-448919-anti-hsp27-antibody-ep1724y-western-blot-hela-lysates.jpg "Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1724Y] to Hsp27
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Hsp27 antibody [EP1724Y]
See all Hsp27 primary antibodies -
Description
Rabbit monoclonal [EP1724Y] to Hsp27 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-Pmore details
Unsuitable for: Flow Cyt or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human Hsp27 (N terminal). The exact sequence is proprietary.
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Positive control
- WB: HeLa, HAP1 and MCF7 cell lysates. IHC-P: Human cervical carcinoma and human breast carcinoma tissues.
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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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1724Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab62339 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 |
|---|---|---|
| WB |
1/500 - 1/2000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa).
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| IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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| Notes |
|---|
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WB
1/500 - 1/2000. Detects a band of approximately 23 kDa (predicted molecular weight: 23 kDa). |
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IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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
- Omim: 602195 Human
- SwissProt: P04792 Human
- Unigene: 520973 Human
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Alternative names
- Heat shock 27kDa protein antibody
- 28 kDa heat shock protein antibody
- CMT2F antibody
see all
Images
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Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)All lanes : Anti-Hsp27 antibody [EP1724Y] (ab62339) at 1/500 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : HSPB1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 23 kDaLanes 1- 2: Merged signal (red and green). Green - ab62339 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab62339 was shown to react with Hsp27 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab261738 (knockout cell lysate ab256945) was used. Wild-type HeLa and HSPB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab62339 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 500 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] (ab62339)](https://www.abcam.com/ps/products/62/ab62339/Images/ab62339-75645-anti-hsp27-antibody-ep1724y-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp27 antibody [EP1724Y] (ab62339)Immunohistochemical analysis of Hsp27 expression in paraffin embedded human cervical carcinoma (left) and human breast carcinoma (right) using 1/100 ab62339
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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![Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)](https://www.abcam.com/ps/products/62/ab62339/Images/ab62339-281777-anti-hsp27-antibody-ep1724y-western-blot.jpg)
Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)All lanes : Anti-Hsp27 antibody [EP1724Y] (ab62339) at 1/500 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : Hsp27 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 23 kDaLanes 1 - 4: Merged signal (red and green). Green - ab62339 observed at 27 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab62339 was shown to specifically react with Hsp27 in wild-type HAP1 cells. No band was observed when Hsp27 knockout samples were used. Wild-type and Hsp27 knockout samples were subjected to SDS-PAGE. Ab62339 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)](https://www.abcam.com/ps/products/62/ab62339/Images/ab62339-75646-anti-hsp27-antibody-ep1724y-western-blot.jpg)
Western blot - Anti-Hsp27 antibody [EP1724Y] (ab62339)Anti-Hsp27 antibody [EP1724Y] (ab62339) at 1/500 dilution + HeLa cell lysate at 10 µg
Secondary
Goat anti-rabbit, HRP labeled at 1/2000 dilution
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted? -
![Anti-Hsp27 antibody [EP1724Y] (ab62339)](https://www.abcam.com/ps/products/62/ab62339/Images/ab62339-5-benefits-of-recombinant-antibodies.png)
Anti-Hsp27 antibody [EP1724Y] (ab62339)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (11)
ab62339 has been referenced in 11 publications.
- Booth L et al. NEDD4 over-expression regulates the afatinib resistant phenotype of NSCLC cells. Oncol Signal 1:19-30 (2018). PubMed: 30740589
- Booth L et al. PDE5 inhibitors enhance the lethality of pemetrexed through inhibition of multiple chaperone proteins and via the actions of cyclic GMP and nitric oxide. Oncotarget 8:1449-1468 (2017). ICC/IF . PubMed: 27903966
- Booth L et al. The HDAC inhibitor AR42 interacts with pazopanib to kill trametinib/dabrafenib-resistant melanoma cells in vitro and in vivo. Oncotarget 8:16367-16386 (2017). PubMed: 28146421
- Booth L et al. PDE5 inhibitors enhance the lethality of [pemetrexed + sorafenib]. Oncotarget 8:13464-13475 (2017). PubMed: 28088782
- Booth L et al. HDAC inhibitors enhance the immunotherapy response of melanoma cells. Oncotarget 8:83155-83170 (2017). PubMed: 29137331