PA3-017

antibody from Invitrogen Antibodies

Targeting: HSF1 HSTF1

Provider product page for PA3-017

Western blot

ELISA

Immunocytochemistry

Immunoprecipitation

Gel shift

Other assay

Antibody data

Antibody Data
Antigen structure
References [26]
Comments [0]
Validations
Immunocytochemistry [1]
Other assay [4]

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Product number: PA3-017 - Provider product page
Provider: Invitrogen Antibodies
Product name: HSF1 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: PA3-017 detects heat shock factor 1 (HSF-1) from human, mouse and rat tissues and cells. PA3-017 has been successfully used in Western blot, immunoprecipitation, immunofluorescence, ELISA, and gel super shift protocols. By Western blot, this antibody detects an ~83 kDa protein representing HSF-1 from NIH-3T3 cell lysate. The PA3-017 immunogen is recombinant human HSF-1 protein expressed in E. coli.
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: Conc. Not Determined
Storage: -20° C, Avoid Freeze/Thaw Cycles

HSF1 protein structure - PA3-017 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunoprecipitation of Heat Shock Factor 1 (HSF 1) was performed on HeLa cells. Antigen:antibody complexes were formed by incubating 500µg whole cell lysate with 2µl of HSF1 polyclonal antibody (Product # PA3-017) overnight on a rocking platform at 4°C. Immune complexes were captured on 50µl Protein A/G Plus Agarose (Product # 20423), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a HSF1 polyclonal antibody (Product # PA3-017) at a dilution of 1:1000 overnight rotating at 4°C, washed in TBST, and probed with Clean Blot IP Detection Reagent-HRP (Product # 21232) at a dilution of 1:1000 for at least one hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 8 Possible mechanisms of the LSC + L treatment in sensitizing the drug-resistant cells to cancer therapy. a Qualitative western blot data of heat shock protein 90 and 70 (HSP90 and HSP70), heat shock factor-1 (HSF-1), and mutant p53 proteins, showing decreased expression of HSP70, HSF-1, and mutant p53 after the treatment of LSC nanoparticles and NIR laser irradiation (LSC + L). b Quantitative data of the western blot results, showing the decreased expression of HSP70, HSF-1, and mutant p53 in the NCI/RES-ADR cells with the LSC + L treatment is statistically significant. Error bars represent s.d. ( n = 3). * p < 0.05 (Kruskal-Wallis H-test). c Flow cytometry analysis showing decreased expression of HSP70, HSF-1, and mutant p53 in NCI/RES-ADR cells treated with LSC nanoparticles and NIR laser irradiation. d Confocal images of HSF-1 showing the HSF-1 distributes in both nuclei and cytoplasm. e Confocal images of HSF70 showing the HSF70 mainly distributes in cytoplasm. f Confocal images of mutant p53 show the mutant p53 mainly distributes in nuclei. The distribution of P-gp is similar to that shown in Fig. 4a . No treatment (laser or nanoparticle) was conducted on the cells in the control group. The cells were permeabilized for the flow cytometry and confocal analyses. The NIR laser irradiation was at 1 W cm -2 for 1 min. The concentration of LSC nanoparticles used for the LSC or LSC + L groups was 0.2 mg ml -1 . Scale bars: 20 mum