HSP27 Monoclonal antibody

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• KD/KO Validated

HSP27 Monoclonal Antibody for WB, IHC, IF, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, zebrafish

Applications

WB, IHC, IF, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

3E7A1

Cat no : 66767-1-Ig

Synonyms

HSPB1, HSP28, Hsp25, HMN2B, Estrogen-regulated 24 kDa protein

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Validation Data Gallery

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  WB analysis using 66767-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66767-1-Ig (HSPB1 antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

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  WB analysis of HeLa using 66767-1-Ig

  WB result of HSP27 antibody (66767-1-Ig; 1:60000; incubated at room temperature for 1.5 hours) with sh-Control and sh-HSP27 transfected HeLa cells.

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  WB analysis of HSC-T6 using 66767-1-Ig

  HSC-T6 cells were subjected to SDS PAGE followed by western blot with 66767-1-Ig (HSPB1 antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

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  WB analysis of mouse liver using 66767-1-Ig

  mouse liver tissue were subjected to SDS PAGE followed by western blot with 66767-1-Ig (HSPB1 antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

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  IHC staining of human gliomas using 66767-1-Ig

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 66767-1-Ig (HSPB1 antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of MCF-7 using 66767-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed MCF-7 cells using HSP27 antibody (66767-1-Ig, Clone: 3E7A1 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

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  IF Staining of zebrafish retina using 66767-1-Ig

  Antibody staining on tissue sections of the zebrafish retina at 5 days post fertilisation (dpf) for Heatshock protein 27 (Hsp27;magenta); (#66767-1-Ig) Dilution 1:200. Tissue fixed overnight in 4% PFA; DAPI, blue. Data generated by Natalia Jaroszynska in Professor Ryan MacDonald's lab, University College London, UK.

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  FC experiment of HepG2 using 66767-1-Ig

  1X10^6 HepG2 cells were intracellularly stained with 0.5 ug Anti-Human HSP27 (66767-1-Ig, Clone:3E7A1) (red) labeled with FlexAble CoraLite® Plus 555 Antibody Labeling Kit for Mouse IgG1 (KFA022), or 0.5 ug Control Antibody. Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: HCT 116 cells, HeLa cells, HSC-T6 cells, mouse liver tissue, U2OS cells, A549 cells, HepG2 cells, HuH-7 cells
• Positive IHC detected in: human gliomas tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF detected in: zebrafish retina, MCF-7 cells
• Positive FC (Intra) detected in: HepG2 cells

Recommended dilution

• Western Blot (WB): WB : 1:20000-1:100000
• Immunohistochemistry (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF): IF : 1:50-1:500
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.50 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below
• IF: See 2 publications below

Product Information

66767-1-Ig targets HSP27 in WB, IHC, IF, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, zebrafish samples.

• Tested Reactivity: human, mouse, rat, zebrafish
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: HSP27 fusion protein Ag27859
• Full Name: heat shock 27kDa protein 1
• Calculated Molecular Weight: 19-23 kDa
• Observed Molecular Weight: 27 kDa
• GenBank Accession Number: BC012768
• Gene Symbol: HSPB1
• Gene ID (NCBI): 3315
• RRID: AB_2882113
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

HSPB1, also known as heat shock protein 27 (Hsp27), belongs to the small heat shock protein family which is induced in response to environmental challenges or/and developmental transitions. It is also an anti-apoptotic protein that plays crucial roles in tumorigenesis and cell survival and is reported to be an independent prognosis marker for cancer. Recently HSPB1 has been found to be a valuable marker for melanoma. In addition to the predicted 27 kDa, an extra 50-55 kDa representing dimeric form of HSPB1 may also be observed (21353161).

Publications

Species	Application	Title
human	IF	Oncol Rep; SUMOylation of HSP27 regulates PKM2 to promote esophageal squamous cell carcinoma progression.; Authors - Xiao Zhang
human,mouse	WB,IF	Sci Adv; Reactive astrocytes secrete the chaperone HSPB1 to mediate neuroprotection; Authors - Fangjia Yang