ET1701-70

Hsp27 Recombinant Rabbit Monoclonal Antibody [JJ09-13]

cat.: ET1701-70

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	JJ09-13

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 peptide within Human Hsp27 aa 127-176 / 205.
Positive control:	Hela cell lysate, A549 cell lysate, Jurkat cell lysate, Hela, HepG2, human tonsil tissue, human colon carcinoma tissue, human breast carcinoma tissue.
Subcellular location:	Cytoplasm, Cytoskeleton, Nucleus.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P FC IP	1:500-1:2,000 1:100-1:500 1:100-1:500 1:50-1:200 1:50-1:100 Use at an assay dependent concentration.
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

	Fig1: Western blot analysis of Hsp27 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 (ET1701-70, 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: Hela cell lysate Lane 2: A549 cell lysate Lane 3: Jurkat cell lysate
	Fig2: ICC staining of Hsp27 in HepG2 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-70, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
	Fig3: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Hsp27 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-70, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig4: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Hsp27 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-70, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Hsp27 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET1701-70, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: Immunocytochemistry analysis of HeLa cells labeling Hsp27 with Rabbit anti-Hsp27 antibody (ET1701-70) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Hsp27 antibody (ET1701-70) 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 (M1305-2, 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.
	Fig7: Flow cytometric analysis of HeLa cells labeling Hsp27. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1701-70, 1μg/mL) (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).

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