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Phospho-Hsp27 (S82) Recombinant Rabbit Monoclonal Antibody [SN06-73]

Usd: 385

Specification

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Catalog# ET1611-16

Phospho-Hsp27 (S82) Recombinant Rabbit Monoclonal Antibody [SN06-73]

Application

  • WB

  • IP

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Phospho-Hsp27 (S82) Recombinant Rabbit Monoclonal Antibody [SN06-73]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser82 of Human Hsp27 aa 50-99 / 205.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IP, IF-Cell, FC

Molecular Weight

Predicted band size: 23 kDa

Positive Control

Human skeletal muscle tissue lysate, MCF-7 cell lysate, MCF7.

Conjugation

unconjugated

Clone Number

SN06-73

RRID

AB_3069993

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:500-1:2,000

  • IP

  • Use at an assay dependent concentration.

  • IF-Cell

  • 1:100

  • FC

  • 1:1,000

Target

Function

HSP 27 is a constitutively expressed cytoplasmic protein that co-localizes to the nucleus upon stress-induced insult. Heat shock, cytokines and hormones are among the factors that stimulate the synthesis of HSP 27. The intracellular concentration of the mammalian heat shock protein, HSP 27, increases several-fold after heat shock and other metabolic stresses, and is closely associated with the acquisition of thermotolerance. MAP kinase-activated protein kinase-2 phosphorylates HSP 27 on Serine residues Ser 15, Ser 78 and Ser 82, which are phosphorylated in vivo in response to growth factors and heat shock. Ser 15, Ser 78 and Ser 82 occur in the sequence motif RXXS, which is recognized by ribosomal protein S6 kinase II.

Background References

1. Leja-Szpak, A. et al. 2015. Kynuramines induce overexpression of heat shock proteins in pancreatic cancer cells via 5-hydroxytryptamine and MT1/MT2 receptors. Journal of physiology and pharmacology : an official journal of the Polish Physiological Society. 66: 711-8.

2. Shreeram, S. et al. 2006. Regulation of ATM/p53-dependent suppression of myc-induced lymphomas by Wip1 phosphatase. J Exp Med. 203(13): 2793-2799.

Sequence Similarity

Belongs to the small heat shock protein (HSP20) family.

Tissue Specificity

Detected in all tissues tested: skeletal muscle, heart, aorta, large intestine, small intestine, stomach, esophagus, bladder, adrenal gland, thyroid, pancreas, testis, adipose tissue, kidney, liver, spleen, cerebral cortex, blood serum and cerebrospinal fluid. Highest levels are found in the heart and in tissues composed of striated and smooth muscle.

Post-translational Modification

Phosphorylated upon exposure to protein kinase C activators and heat shock. Phosphorylation by MAPKAPK2 and MAPKAPK3 in response to stress dissociates HSPB1 from large small heat-shock protein (sHsps) oligomers and impairs its chaperone activity and ability to protect against oxidative stress effectively. Phosphorylation by MAPKAPK5 in response to PKA stimulation induces F-actin rearrangement.

Subcellular Location

Cytoplasm, Nucleus.

UNIPROT

SwissProt: P04792 Human

SwissProt: P14602 Mouse

SwissProt: P42930 Rat

Synonyms

Heat shock 27kDa protein antibody

28 kDa heat shock protein antibody

CMT2F antibody

DKFZp586P1322 antibody

epididymis secretory protein Li 102 antibody

Estrogen regulated 24 kDa protein antibody

Estrogen-regulated 24 kDa protein antibody

Heat shock 25kDa protein 1 antibody

Heat shock 27 kDa protein antibody

Heat shock 27kD protein 1 antibody

Expand

Images

  • Western blot analysis of Phospho-Hsp27 (S82) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1611-16" style="font-weight: bold;text-decoration: underline;">ET1611-16</a>, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:5,000 dilution was used for 1 hour at room temperature.<br /><span style="font-weight: bold;">Positive control:</span> <br />Lane 1: human skeletal muscle tissue lysate<br />Lane 2: MCF-7 cell lysate

    Western blot analysis of Phospho-Hsp27 (S82) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1611-16, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
    Positive control:
    Lane 1: human skeletal muscle tissue lysate
    Lane 2: MCF-7 cell lysate

  • Immunocytochemistry analysis of MCF7 cells labeling Phospho-Hsp27 (S82) with Rabbit anti-Phospho-Hsp27 (S82) antibody (<a href="/products/ET1611-16" style="font-weight: bold;text-decoration: underline;">ET1611-16</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Hsp27 (S82) antibody (<a href="/products/ET1611-16" style="font-weight: bold;text-decoration: underline;">ET1611-16</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of MCF7 cells labeling Phospho-Hsp27 (S82) with Rabbit anti-Phospho-Hsp27 (S82) antibody (ET1611-16) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-Hsp27 (S82) antibody (ET1611-16) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of MCF7 cells labeling Phospho-Hsp27 (S82).<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1611-16" style="font-weight: bold;text-decoration: underline;">ET1611-16</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of MCF7 cells labeling Phospho-Hsp27 (S82).

    Cells were fixed and permeabilized. Then stained with the primary antibody (ET1611-16, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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