Phospho-Hsp27 (S82) Recombinant Rabbit Monoclonal Antibody [SN06-73]
cat.: ET1611-16
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IP, IF-Cell, FC
Clonality: Monoclonal
Clone number: SN06-73
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 23 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser82 of Human Hsp27 aa 50-99 / 205.
Positive control: Human skeletal muscle tissue lysate, MCF-7 cell lysate, MCF7.
Subcellular location: Cytoplasm, Nucleus.
Recommended Dilutions:
  WB
  IP
  IF-Cell
  FC

1:500-1:2,000
Use at an assay dependent concentration.
1:100
1:1,000
Uniprot #: SwissProt: P04792 Human | P14602 Mouse | P42930 Rat
Alternative names: Heat shock 27kDa protein 28 kDa heat shock protein CMT2F DKFZp586P1322 epididymis secretory protein Li 102 Estrogen regulated 24 kDa protein Estrogen-regulated 24 kDa protein Heat shock 25kDa protein 1 Heat shock 27 kDa protein Heat shock 27kD protein 1 Heat shock 27kDa protein 1 Heat shock 28kDa protein 1 Heat Shock Protein 27 Heat shock protein beta 1 Heat shock protein beta-1 heat shock protein family B (small) member 1 HEL-S-102 HMN2B HS.76067 Hsp 25 HSP 27 Hsp 28 Hsp B1 Hsp25 HSP27 Hsp28 HspB1 HSPB1_HUMAN SRP27 Stress responsive protein 27 Stress-responsive protein 27
Images
ET1611-16_1.jpg Fig1: 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
ET1611-16_2.jpg Fig2: 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.
ET1611-16_3.jpg Fig3: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.