Phospho-HSP27 (Ser15) Recombinant antibody

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  WB analysis of HeLa using 83332-3-RR

  Non-treated HeLa cells, and anisomycin treated HeLa cells were subjected to SDS PAGE followed by western blot with 83332-3-RR (Phospho-HSP27 (Ser15) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Beta Actin(66009-1-Ig) antibody as loading control.

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  IF Staining of HeLa using 83332-3-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed Anisomycin treated HeLa cells using Phospho-HSP27 (Ser15) antibody (83332-3-RR, Clone: 2D4 ) at dilution of 1:400 and CoraLite®488-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HeLa using 83332-3-RR

  1X10^6 HeLa cells untreated (dashed lines) or treated with λ phosphatase which intracellularly stained with 0.13 ug Phospho-HSP27 (Ser15) Recombinant antibody (83332-3-RR, Clone:2D4) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.13 ug Rabbit IgG Isotype Control RecAb (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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Phospho-HSP27 (Ser15) Uni-rAb^TM Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA

Cat No. 83332-3-RR

Clone No.2D4

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IF/ICC, FC (Intra), ELISA

Synonyms

Hsp27 (phospho S15), HSP27 (Ser15), p HSP27 , p HSP27 (Ser15), p HSP27 Ser15

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Tested Applications

• Positive WB detected in: Anisomycin treated HeLa cells
• Positive IF/ICC detected in: Anisomycin treated HeLa cells
• Positive FC (Intra) detected in: λ phosphatase treated HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.13 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.

Product Information

83332-3-RR targets Phospho-HSP27 (Ser15) in WB, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: Peptide
• 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_3670996
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• UNIPROT ID: P04792
• 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 named as HSP27, belongs to the mammalian small HSP family widely expressed in cells including the cytoplasm and nucleus. It has been reported that HSPB1 plays a key role in stress resistance, actin organization and numerous biological processes. HSPB1 can be phosphorylated in different stimulation such as cytokines, growth factors and peptide hormones on the site of Ser15, Ser78 and Ser82 by MAPKAP2 kinase. The phosphorylation of HSPB1 leads to a decrease in the size of HSPB1 oligomers. HSPB1 is associated with amyotrophic lateral sclerosis (ALS) (PMID: 8542580, 11960785, 10383393, 32231288)