AB281198

Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture)

KO Validated
RabMAb
Recombinant
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Knockout Tested Rabbit Recombinant Monoclonal Hsp27 antibody. Carrier free. Suitable for WB, ICC/IF, sELISA and reacts with Human samples.

View Alternative Names

HSP27, HSP28, HSPB1, Heat shock protein beta-1, HspB1, 28 kDa heat shock protein, Estrogen-regulated 24 kDa protein, Heat shock 27 kDa protein, Heat shock protein family B member 1, Stress-responsive protein 27, HSP 27, SRP27

4 Images

Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Immunocytochemistry/ Immunofluorescence analysis of wild-type and HSPB1 KO HAP1 (human chronic myelogenous leukemia near-haploid cell) labeling Hsp27 with ab281198 at 1/100 dilution followed by ab150081, secondary antibody at 1/1000 dilution. Confocal image showing cytoplasmic staining in wild-type HAP1 cells, and showing no staining in HSPB1 knockout HAP1 cells (shown in green). The counterstain antibody was ab195889 at 1/200 and was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% TritonX-100. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Sandwich ELISA - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

sELISA

Supplier Data

Sandwich ELISA - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Sandwich ELISA of ab308542 with the capture antibody dilution at 2 µg/mL and detector antibody dilution at 0.5 µg/mL. A549 + Anisomycin : A549 treated with 1 µg/ml anisomycin for 30 minutes, whole cell lysate. Serial dilutions of lysates were analyzed. A549 Control : untreated A549, whole cell lysate. Serial dilutions of lysates were analyzed.

Lab

Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Loading control : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1/200000) (36KDa). Blocking/Diluting buffer and concentration 5% NFDM/TBST Negative control : HT-1080.

All lanes:

Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 27 kDa

false

Exposure time: 180s

Lab

Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Loading control : Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1/200000) (36KDa). Blocking/Diluting buffer and concentration 5% NFDM/TBST Low expression : cerebellum and spleen

All lanes:

Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human liver tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 23-27 kDa

false

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR23938-256

Isotype

IgG

Carrier free

Yes

Reacts with

Human

Applications

ICC/IF, WB, sELISA

applications

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "sELISA" : {"fullname" : "Sandwich ELISA", "shortname":"sELISA"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/1000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/100", "ICCIF-species-notes": "", "sELISA-species-checked": "guaranteed", "sELISA-species-dilution-info": "", "sELISA-species-notes": "" } } }

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Product details

Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (ab281198) is a BSA and Azide Free antibody supplied in an unconjugated format and it is suitable for sandwich ELISAs to quantify human Hsp27.

The recommended pair for sandwich ELISA is:

Capture: ab281198, human Hsp27 Capture Antibody (unconjugated)

Detector: ab308543, human Hsp27 Detector Antibody (unconjugated)

The recommended antibody orientation is based on internal optimization for ELISA-based assays. Antibody orientation is assay dependent and needs to be optimized for each assay type. Please note that the range provided for this antibody is only an estimation based on the performance of the product using the recommended antibody pair. Performance of the antibody pair will depend on the specific characteristics of your assay. We guarantee the product works in sandwich ELISA, but we do not guarantee the sensitivity or dynamic range of the antibody in your assay.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency. The antibodies are provided at an approximate concentration of 1 mg/ml as measured by the protein A280 method. 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.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. 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.

Sandwich ELISA
The recommended antibody orientation is based on internal optimization for ELISA-based assays. Antibody orientation is assay dependent and needs to be optimized for each assay type. Please note that the range provided for this antibody is only an estimation based on the performance of the product using the recommended antibody pair. Performance of the antibody pair will depend on the specific characteristics of your assay. We guarantee the product works in sandwich ELISA, but we do not guarantee the sensitivity or dynamic range of the antibody in your assay.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:

Visit the General protocols
Visit the Troubleshooting

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Target data

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).

See full target information HSPB1

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture)

Immunocytochemistry/ Immunofluorescence - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Sandwich ELISA - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Western blot - Anti-Hsp27 antibody [EPR23938-256] - BSA and Azide free (Capture) (AB281198)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Complementary Products

Assay Kits

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Product promise