Rabbit Recombinant Monoclonal Hsp27 antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with African green monkey, Human samples.
Images
![Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--western-blot-img136698.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442)")
![Flow Cytometry (Intracellular) - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--flow-cytometry-intracellular-img63850.jpg/_jcr_content/renditions/webp-475.webp "Flow Cytometry (Intracellular) - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442)")
![Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--immunocytochemistry-immunofluorescence-img68865.jpg/_jcr_content/renditions/webp-475.webp "Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442)")
![Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--western-blot-img100243.jpg/_jcr_content/renditions/webp-475.webp "Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (AB229442)")
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
Liquid
- Clonality
Monoclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
Reactivity data
Select an application| WB | ICC/IF | Flow Cyt (Intra) | |
|---|---|---|---|
| Human | Tested | Tested | Tested |
| Rat | Predicted | Predicted | Predicted |
| African green monkey | Tested | Expected | Expected |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species African green monkey, Human | Dilution info - | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes - |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species African green monkey | Dilution info Use at an assay dependent concentration. | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info - | Notes - |
Flow Cyt (Intra)
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info - | Notes ab199376 -Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species African green monkey | Dilution info - | Notes ab199376 -Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody. |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Rat | Dilution info - | Notes - |
Associated Products
Select an associated product type
15 products for Alternative Product
3 products for Alternative Version
Target data
Function
Small heat shock protein which functions as a molecular chaperone probably maintaining denatured proteins in a folding-competent state (PubMed:10383393, PubMed:20178975). Plays a role in stress resistance and actin organization (PubMed:19166925). Through its molecular chaperone activity may regulate numerous biological processes including the phosphorylation and the axonal transport of neurofilament proteins (PubMed:23728742).
Alternative names
Heat shock protein beta-1, HspB1, 28 kDa heat shock protein, Estrogen-regulated 24 kDa protein, Heat shock 27 kDa protein, Stress-responsive protein 27, HSP 27, SRP27, HSP28, HSP27, HSPB1
Recommended products
Rabbit Recombinant Monoclonal Hsp27 antibody. Carrier free. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with African green monkey, Human samples.
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.2 - 7.4
Constituents: PBS- Form
Liquid
- Clonality
Monoclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Carrier free
Yes
- Clone number
EPR5477
- Purification technique
Affinity purification Protein A
- Concentration
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
+4°C
- Storage information
Do Not Freeze
Notes
ab229442 is the carrier-free version of Anti-Hsp27 antibody [EPR5477] ab109376.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Hsp27 also known as HSPB1 is a small heat shock protein with a molecular weight of approximately 27 kilodaltons. This protein is expressed in various tissues including muscle heart and brain. It functions as a molecular chaperone that stabilizes unfolded proteins preventing their aggregation. Hsp27 undergoes phosphorylation at specific residues which modulates its chaperone activity and interaction with other proteins.
- Biological function summary
Hsp27 plays a critical role in cellular stress response by regulating actin cytoskeleton dynamics and inhibiting apoptosis. It forms part of a complex that includes other proteins such as alphaB-crystallin. This complex facilitates the reorganization of proteins under stress conditions enhancing cell survival during oxidative stress or thermal shock. Hsp27 also modulates inflammatory responses and has been shown to affect cell migration.
- Pathways
Hsp27 integrates into the apoptosis and inflammation pathways. It interacts with apoptotic machinery such as caspase proteins to protect cells by hindering apoptosome formation. Additionally Hsp27 can engage with pathways involving the nuclear factor-kappa B (NF-kB) impacting inflammatory signaling. CPTC (carboxyl-pyrene-trioctylamine) can modulate these pathways by altering Hsp27 function and interactions.
- Associated diseases and disorders
Hsp27 has connections to neurodegenerative diseases and cancer. In neurodegenerative conditions such as Alzheimer's disease its chaperone activity is thought to protect neurons from misfolded protein aggregates. In cancer Hsp27 supports tumor cell survival and resistance to chemotherapy by interacting with proteins like Akt and p53. These interactions highlight the complex role of Hsp27 in modulating cellular responses in various pathological states.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
4 product images
![Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--western-blot-img136698.jpg "Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)")
Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)
This data was developed using the same antibody clone in a different buffer formulation (Anti-Hsp27 antibody [EPR5477] ab109376).
Lanes 1- 2: Merged signal (red and green). Green - Anti-Hsp27 antibody [EPR5477] ab109376 observed at 23 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
Anti-Hsp27 antibody [EPR5477] ab109376 was shown to react with Hsp27 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human HSPB1 (Hsp27) knockout HeLa cell line ab261738 (knockout cell lysate Human HSPB1 (Hsp27) knockout HeLa 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. Anti-Hsp27 antibody [EPR5477] ab109376 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (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.
All lanes: Western blot - Anti-Hsp27 antibody [EPR5477] (Anti-Hsp27 antibody [EPR5477] ab109376) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: HSPB1 knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 23 kDa
![Flow Cytometry (Intracellular) - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--flow-cytometry-intracellular-img63850.jpg "Flow Cytometry (Intracellular) - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)")
Flow Cytometry (Intracellular) - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)
Overlay histogram showing HAP1 wildtype (green line) and HAP1-HSPB1 knockout cells (red line) stained with Anti-Hsp27 antibody [EPR5477] 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 (Anti-Hsp27 antibody [EPR5477] ab109376, 1�g/ml) for 30 min at 22�C. The secondary antibody used was Alexa Fluor� 488 goat anti-rabbit IgG (H&L) presorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22�C. A rabbit IgG isotype control antibody� (Rabbit IgG, monoclonal [EPR25A] - Isotype Control 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Hsp27 antibody [EPR5477] ab109376).
![Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--immunocytochemistry-immunofluorescence-img68865.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)")
Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)
This ICC/IF data was generated using the same anti-Hsp27 antibody clone, EPR5477, in a different buffer formulation (cat# Anti-Hsp27 antibody [EPR5477] ab109376).
Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Hsp27 with purified Anti-Hsp27 antibody [EPR5477] ab109376 at 1/500. Cells were fixed with 100% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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.
![Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442), expandable thumbnail](https://content.abcam.com/products/images/hsp27-antibody-epr5477-bsa-and-azide-free-ab229442--western-blot-img100243.jpg "Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)")
Western blot - Anti-Hsp27 antibody [EPR5477] - BSA and Azide free (ab229442)
This WB data was generated using the same anti-Hsp27 antibody clone, EPR5477, in a different buffer formulation (cat# Anti-Hsp27 antibody [EPR5477] ab109376).
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Hsp27 antibody [EPR5477] ab109376 observed at 27 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-Hsp27 antibody [EPR5477] 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. Anti-Hsp27 antibody [EPR5477] ab109376 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Hsp27 antibody [EPR5477] (Anti-Hsp27 antibody [EPR5477] ab109376) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: Hsp27 knockout HAP1 whole cell lysate at 20 µg
Lane 3: HeLa whole cell lysate at 20 µg
Lane 4: MCF7 whole cell lysate at 20 µg
Predicted band size: 23 kDa
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