phospho-HSF1 (Ser326) Recombinant Antibody

$MCF7 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HSF1 (S326) (37E4) Monoclonal Antibody(bsm-52166R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).$

MCF7 cells(black) were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20\u2103, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HSF1 (S326) (37E4) Monoclonal Antibody(bsm-52166R)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody(blue) incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

$Paraformaldehyde-fixed, paraffin embedded Human endometrial carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with HSF1 (S326) (37E4) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human endometrial carcinoma; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with HSF1 (S326) (37E4) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, DAB staining.

$Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HSF1 (S326)) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.$

Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with (HSF1 (S326)) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

$HeLa (H) treated with heat at 43\u2103 for 30 min, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-HSF1 (Ser326) monoclonal antibody, unconjugated (bsm-52166R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

HeLa (H) treated with heat at 43\u2103 for 30 min, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-HSF1 (Ser326) monoclonal antibody, unconjugated (bsm-52166R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$The HeLa (treated with heat (44\u00baC)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature\uff0cfollowed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-HSF1 (Ser326) antibody (bsm-52166R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.$

The HeLa (treated with heat (44\u00baC)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.). Cells stained with Primary Antibody for 30 min at room temperature\uff0cfollowed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-phospho-HSF1 (Ser326) antibody (bsm-52166R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.

$Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Ovarian Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Prostate Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Prostate Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with phospho-HSF1 (Ser326) Monoclonal Antibody, Unconjugated (bsm-52166R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.

Applications

WB
IHC-P
IHC-F
IF
Flow-Cyt

Reactivity

Human

Predicted Reactivity

Overview
Catalog #	bsm-52166r
Product Name	phospho-HSF1 (Ser326) Recombinant Antibody
Applications	WB IHC-P IHC-F IF FLOW-CYT
Reactivity	Human
Predicted Reactivity	Rat
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	KLH conjugated Synthesised phosphopeptide derived from human HSF1 around the phosphorylation site of Ser326
Modification Site	S326
Clonality	Recombinant
Clone #	37E4
Isotype	IgG
Concentration	1mg/ml
Purification	purified by Protein A.
Storage Buffer	0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID	3297
Swiss Prot	Q00613
Subcellular location	Cytoplasm, Nucleus
Synonyms	HSTF1; HSTF; HSTF 1; Hsf1alpha; Hsf1beta; HSF1_HUMAN; HSF1; HSF 1; Heat shock transcription factor 1 (HSTF 1); HSF1_MOUSE; heat shock transcription factor 1; HSF1 (phospho-S326); p-HSF1; phospho-HSF1; HSF1 (phospho-Ser326)
Background	DNA-binding protein that specifically binds heat shock promoter elements (HSE) and activates transcription. In higher eukaryotes, HSF is unable to bind to the HSE unless the cells are heat shocked.
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IHC-F	1:100-500
IF
Flow-Cyt	1ug/Test