Product nameAnti-Hsp27 antibody [EPR5477]
See all Hsp27 primary antibodies
DescriptionRabbit monoclonal [EPR5477] to Hsp27
Tested applicationsSuitable for: ICC/IF, WB, IP, IHC-P, Flow Cytmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human Hsp27 aa 100-200. The exact sequence is proprietary.
(Peptide available as
- HeLa, HAP1, COS-1, BxPC-3 and HT-1376 cell lysates Flow Cyt: HAP1-WT cells.
Storage instructionsShipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
Immunizing Peptide (Blocking)
Our Abpromise guarantee covers the use of ab109376 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/10000. Predicted molecular weight: 23 kDa.Can be blocked with Hsp27 peptide (ab215449).|
|IP||1/10 - 1/100.|
|IHC-P||1/100 - 1/250. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.|
|Flow Cyt||1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
FunctionInvolved in stress resistance and actin organization.
Tissue specificityDetected 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 diseaseDefects 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 similaritiesBelongs to the small heat shock protein (HSP20) family.
modificationsPhosphorylated in MCF-7 cells on exposure to protein kinase C activators and heat shock.
Cellular localizationCytoplasm. 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
- Heat shock 27kDa protein antibody
- 28 kDa heat shock protein antibody
- CMT2F antibody
Lane 1: Wild type HAP1 whole cell lysate (20 µg)
Lane 2: Hsp27 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: MCF7 whole cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab109376 observed at 27 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab109376 was shown to specifically react with Hsp27 in wild-type cells as signal was lost in Hsp27 knockout HEP1 cells. Wild-type and Hsp27 knockout samples were subjected to SDS-PAGE. Ab109376 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 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.
Overlay histogram showing HAP1 wildtype (green line) and HAP1-HSPB1 knockout cells (red line) stained with ab109376. The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab109376, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C.
A rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-HSPB1 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Hsp27 with purified ab109376 at 1/500. Cells were fixed with 100% methanol. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).
Secondary Only Control: PBS was used instead of the primary antibody as the negative control.
All lanes : Anti-Hsp27 antibody [EPR5477] (ab109376) at 1/1000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : COS-1 cell lysate
Lane 3 : BxPC-3 cell lysate
Lane 4 : HT-1376 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 23 kDa
Observed band size: 27 kDa why is the actual band size different from the predicted?
This product has been referenced in:
- Tanaka T et al. Sec6 enhances cell migration and suppresses apoptosis by elevating the phosphorylation of p38 MAPK, MK2, and HSP27. Cell Signal 49:1-16 (2018). IP . Read more (PubMed: 29729335) »
- Qiao Y et al. High Glucose Stimulates Tumorigenesis in Hepatocellular Carcinoma Cells Through AGER-Dependent O-GlcNAcylation of c-Jun. Diabetes 65:619-32 (2016). Read more (PubMed: 26825459) »