HSP27 Antibody

Knockout Validated: HSP27 Antibody [NBP1-75477] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout (KO) HeLa cell line. PVDF membrane was probed with 2 ug/ml of Rabbit Anti-Human HSP27 Polyclonal Antibody (Catalog # NBP1-75477) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (catalog number HAF008). Specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.

Western Blot: Hsp27 Antibody [NBP1-75477] - analysis of Jurkat cell lysate (30ug) using anti-Hsp27 antibody. Image from verified customer review.

Immunocytochemistry/Immunofluorescence: Hsp27 Antibody [NBP1-75477] - HSP27 antibody was tested in HeLa cells with FITC (green). Nuclei and actin were counterstained with Dapi (blue) and Phalloidin (red).

Immunohistochemistry: Hsp27 Antibody [NBP1-75477] - Staining of HSP27 in mouse heart using DAB with hematoxylin counterstain.

Western Blot: Hsp27 Antibody [NBP1-75477] - Analysis of HSP27 in human skeletal muscle lysate.

• Reactivity: Hu, Mu, Rt, Ca, Ha
• Applications: WB, ICC/IF, IHC, IHC-P, KO
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 1 mg/ml

HSP27 Antibody Summary

• Immunogen: A synthetic peptide made to an internal region of the human Hsp27 protein (within residues 80-150). [Swiss-Prot P04792]
• Localization: Cytoplasm and nucleus. Translocates to the nucleus during heat shock and resides in sub-nuclear structures known as SC35 speckles or nuclear splicing speckles.
• Predicted Species: Hamster (94%), Canine (94%). Backed by our 100% Guarantee.
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HSPB1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/ml
  • Immunocytochemistry/Immunofluorescence 1:100
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Paraffin 1:100
  • Knockout Validated 2 ug/ml
• Application Notes: This HSP27 antibody is useful for ICC/IF, IHC-P, and Western blot where a band is seen ~27 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.
• Positive Control:

  Skeletal Muscle Lysate (NB820-59253)
  HSP27 Lysate (NBP2-64653)

Reactivity Notes

Human, mouse and rat. Immunogen has 94% identity to canine and hamster.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS and 30% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for HSP27 Antibody

• 28 kDa heat shock protein
• DKFZp586P1322
• Estrogen-regulated 24 kDa protein
• Heat shock 27 kDa protein
• heat shock 27kD protein 1
• heat shock 27kDa protein 1
• heat shock protein beta-1
• HMN2B
• HS.76067
• HSP25
• HSP27
• HSP27HSP 27
• HSP28CMT2F
• HSPB1
• SRP27
• Stress-responsive protein 27

Background

Heat shock protein 27 is a molecular chaperone involved in protection of cells during cellular stress events. The activity of HSP27 protects from apoptotic events caused by heat shock and other distress. Immunohistochemical detection of HSP27 has been suggested as a qualitative biomarker for colorectal cancer. It is also highly expressed in lung cancer, cerebral inflammation, ischemia-reperfusion injury, and glaucoma. Interestingly, HSP27 overexpression can counteract the activity of multi-drug resistance proteins in breast cancer. Polyphenols downregulate the expression of HSP27, as well as many antioxidant enzymes, resulting in susceptibility to stress and damage.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for HSP27 Antibody (NBP1-75477)(2)

Publications using NBP1-75477

• Lopez-Gonzalez I, Carmona M, Arregui L et al. aB-crystallin and HSP27 in glial cells in tauopathies. Neuropathology 2014 Jul 02 [PMID: 24985029] (IHC-P)
  
  Details:
  Hsp27 antibody used for IHC-P for immunostaining HSP27 in glial cells with /without tau deposits in progressive supranuclear palsy, corticobasal degeneration (CBD), argyrophilic grain disease (AGD), Pick's disease (PiD), Alzheimer's disease, frontotemporal lobar degeneration associated with mutations in the tau gene (FTLD-tau), globular glial tauopathy (GGT) and tauopathy in the elderly. Sections boiled in citrate buffer for 20 min to retrieve tau antigenicity, blocked with 1% H2O2 in methanol followed by 3% normal horse serum 1:100 incubated at 4C ON followed by peroxidase-DAB detection with EnVision kit. Antibody also used for double-labeling immunofluorescence and confocal microscopy of paraffin tissue sections.
• Stice JP, Chen L, Kim SC et al. 17Beta-Estradiol, aging, inflammation, and the stress response in the female heart Endocrinology 2011 Apr [PMID: 21303943] (WB, Rat)

Review for HSP27 Antibody (NBP1-75477) (1)

Average Rating: 3

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Anonymous WB Human 10/07/2014

Summary

• Application: Western Blot
• Sample Tested: Whole cell lysate from Jurkat cells, in lysis buffer
• Species: Human
• Comments: The antibody gives a faint band after a very long exposure, which is the case with most other HSP27 antibodies, because of the low levels of HSPs in cells

Blocking

• Blocking Details: 5% milk in TBST buffer

Primary Anitbody

• Dilution Ratio: 1:1000, diluted in TBST buffer+5% milk

Secondary Antibody

• Secondary Description: goat anti-rabbit IgG-HRP (SCBT)
• Secondary Manufacturer Cat#: sc-2004
• Secondary Concentration: 1:10000

Details

• Detection Notes: Detected using Santa Cruz Western Blotting Luminol Reagent, Exposure 2 min

Comments

• Comments: The antibody gives a faint band after a very long exposure, which is the case with most other HSP27 antibodies, because of the low levels of HSPs in cells

Bioinformatics

• Gene Symbol: HSPB1
• Entrez:
  • 3315(Human)
  • 15507(Mouse)
  • 15510(Mouse)
  • 24471(Rat)
• Uniprot:
  • P04792()
  • P14602(Mouse)