Anti-HSF1 Antibody [4B4]

Rat Anti-Mouse HSF1 Monoclonal IgG1

Catalog No. SMC-477

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Species Reactivity Hu, Ms, Rt, Bv, GP, Hm, Mk, Rb
Applications WB IHC ICC/IF FCM IP
SKU: SMC-477 Categories: ,

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SMC-477_HSF1_Antibody_4B4_ICC-IF_Human_Heat-Shocked-HeLa-Cells_100x_Composite.png
Rat Anti-HSF1 Antibody [4B4] used in Immunohistochemistry (IHC) on Human Breast carcinoma (SMC-477)Rat Anti-HSF1 Antibody [4B4] used in Western Blot (WB) on Human Breast adenocarcinoma cell line (MCF7) (SMC-477)Rat Anti-HSF1 Antibody [4B4] used in Immunohistochemistry (IHC) on Mouse Lung (SMC-477)Rat Anti-HSF1 Antibody [4B4] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human Heat Shocked cervical cancer cells (HeLa) (SMC-477)
Product Name HSF1 Antibody
Description

Rat Anti-Mouse HSF1 Monoclonal IgG1
Species Reactivity Bovine, Guinea Pig (Cavia porcellus), Hamster, Human, Monkey, Mouse, Rabbit, Rat
Applications WB, IHC, ICC/IF, IP, ELISA, GS
Antibody Dilution WB (1:1000), IHC (1:2000), ICC/IF (1:100); optimal dilutions for assays should be determined by the user.
Host Species Rat
Immunogen Species Mouse
Immunogen Purified recombinant mouse HSF1 protein, epitope mapping to amino acids 425-439
Concentration 1 mg/ml
Conjugates APC, ATTO 390, ATTO 488, ATTO 594, Biotin, FITC, HRP, PerCP, RPE, Unconjugated
Dylight 488
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1011 g/mol

Dylight 488 Datasheet

 Dylight 488 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 493 nm

λem = 518 nm

εmax = 7.0×104

Laser = 488 nm

 

APC/Cy7
Overview:

  • High quantum yield
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested

APC-Cy7 Datasheet

 

 ACP-Cy7 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 652 nm

λem = 790 nm

Laser = 594 or 633 nm

 

 

  Dylight 350
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent solubility in water
  • Stringently QC tested
  • Excellent batch-to-batch reproducibility
  • Molecular weight: 874 g/mol

Dylight 350 Datasheet

 Dylight 350 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 353 nm

λem = 432 nm

εmax = 1.5×104

 

 

  Dylight 405
Overview:

  • High fluorescence intensity
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 793 g/mol

Dylight 405 Datasheet

 Dylight 405 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 400 nm

λem = 420 nm

εmax = 3.0×104

Laser = 405 nm

 

Dylight 594
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1078 g/mol

Dylight 594 Datasheet

 Dylight 594 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 593 nm

λem = 618 nm

εmax = 8.0×104

Laser = 526 nm

 

 Dylight 633
Overview:

  • High fluorescence yield
  • High photostability
  • Less pH-sensitive
  • Excellent batch-to-batch reproducibility
  • Stringently QC tested
  • Molecular weight: 1066 g/mol

Dylight 633 Datasheet

 Dylight 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 638 nm

λem = 658 nm

εmax = 1.7×105

Laser = 633 nm

 

 PerCP 
Overview:

  • Peridinin-Chlorophyll-Protein Complex
  • Small phycobiliprotein
  • Isolated from red algae
  • Large stokes shift (195 nm)
  • Molecular Weight: 35 kDa
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

PerCP Datasheet

  PerCP Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 482 nm

λem = 677 nm

εmax = 1.96 x 106

Laser = 488 nm

 

 PE/ATTO 594
PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation

PE/ATTO 594 Datasheet

 PE-ATTO 594 Fluorophore Conjugate Excitation and Emission Spectra Optical Properties:

λex = 535 nm

λem = 627 nm

Laser = 488 to 561 nm

 

  FITC (Fluorescein)
Overview:

  • Excellent fluorescence quantum yield
  • High rate of photobleaching
  • Good solubility in water
  • Broad emission spectrum
  • pH dependent spectra
  • Molecular formula: C20H12O5
  • Molar mass: 332.3 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

FITC-Fluorescent-conjugate

 FITC Fluorescein Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 494 nm

λem = 520 nm

εmax = 7.3×104

Φf = 0.92

τfl = 5.0 ns

Brightness = 67.2

Laser = 488 nm

Filter set = FITC

 

 ATTO 700
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 575 g/mol

ATTO 700 Datasheet

  ATTO 700 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 700 nm

λem = 719 nm

εmax = 1.25×105

Φf = 0.25

τfl = 1.6 ns

Brightness = 31.3

Laser = 676 nm

Filter set = Cy®5.5

 

 ATTO 680
Overview:

  • High fluorescence yield
  • Excellent thermal and photostability
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 631 g/mol

ATTO 680 Datasheet

  ATTO 680 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 680 nm

λem = 700 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.7 ns

Brightness = 37.5

Laser = 633 – 676 nm

Filter set = Cy®5.5

 

 ATTO 655
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Excellent ozone resistance
  • Quenched by electron donors
  • Very hydrophilic
  • Good solubility in polar solvents
  • Zwitterionic dye
  • Molar Mass: 634 g/mol

ATTO 655 Datasheet

 ATTO 655 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 663 nm

λem = 684 nm

εmax = 1.25×105

Φf = 0.30

τfl = 1.8 ns

Brightness = 37.5

Laser = 633 – 647 nm

Filter set = Cy®5

 

 ATTO 633
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Stable at pH 4 – 11
  • Cationic dye, perchlorate salt
  • Molar Mass: 652.2 g/mol

ATTO 633 Datasheet

 ATTO 633 Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 629 nm

λem = 657 nm

εmax = 1.3×105

Φf = 0.64

τfl = 3.2 ns

Brightness = 83.2

Laser = 633 nm

Filter set = Cy®5

 

 ATTO 594
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 1137 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

ATTO 594 Datasheet

  ATTO 594 Fluorophore Excitation and Emission Spectrum Optical Properties:

λex = 601 nm

λem = 627 nm

εmax = 1.2×105

Φf = 0.85

τfl = 3.5 ns

Brightness = 102

Laser = 594 nm

Filter set = Texas Red®

 

 ATTO 565
Overview:

  • High fluorescence yield
  • High thermal and photostability
  • Good solubility in polar solvents
  • Excellent solubility in water
  • Very little aggregation
  • Rhodamine dye derivative
  • Molar Mass: 611 g/mol

ATTO 565 Datasheet

  ATTO 565 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 563 nm

λem = 592 nm

εmax = 1.2×105

Φf = 0.9

τfl = 3.4 n

Brightness = 10

Laser = 532 nm

Filter set = TRITC

 

  ATTO 488
Overview:

  • High fluorescence yield
  • High photostability
  • Very hydrophilic
  • Excellent solubility in water
  • Very little aggregation
  • New dye with net charge of -1
  • Molar Mass: 804 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications. 

ATTO 488 Datasheet

   ATTO 488 Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 501 nm

λem = 523 nm

εmax = 9.0×104

Φf = 0.80

τfl = 4.1 ns

Brightness = 72

Laser = 488 nm

Filter set = FITC

 

  ATTO 390
Overview:

  • High fluorescence yield
  • Large Stokes-shift (89 nm)
  • Good photostability
  • Moderately hydrophilic
  • Good solubility in polar solvents
  • Coumarin derivate, uncharged
  • Low molar mass: 343.42 g/mol
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications. 

ATTO 390 Datasheet

 ATTO 390 Fluorescent Dye Excitation and Emission Spectra Optical Properties:

λex = 390 nm

λem = 479 nm

εmax = 2.4×104

Φf = 0.90

τfl = 5.0 ns

Brightness = 21.6

Laser = 365 or 405 nm

 

APC (Allophycocyanin)
Overview:

  • High quantum yield
  • Large phycobiliprotein
  • 6 chromophores per molecule
  • Isolated from red algae
  • Molecular Weight: 105 kDa
  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

APC Datasheet

  APC Fluorophore Absorption and Emission Spectrum Optical Properties:

λex = 650 nm

λem = 660 nm

εmax = 7.0×105

Φf = 0.68

Brightness = 476

Laser = 594 or 633 nm

Filter set = Cy®5

 

Streptavidin

Properties:

  • Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinity
  • Molecular weight: 53 kDa
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA

Streptavidin Datasheet

BiotinBiotin Conjugate Structure

Properties:

  • Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinity
  • Molar mass: 244.31 g/mol
  • Formula: C10H16N2O3S
  • Applications: Western blot, immunohistochemistry, and ELISA*

*The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

Biotin Datasheet

HRP (Horseradish peroxidase)

Properties:

  • Enzymatic activity is used to amplify weak signals and increase visibility of a target
  • Readily combines with hydrogen peroxide (H2O2) to form HRP-H2O2 complex which can oxidize various hydrogen donors
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured products
    • Chemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)
    • Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent products
  • High turnover rate enables rapid generation of a strong signal
  • 44 kDa glycoprotein
  • Extinction coefficient: 100 (403 nm)
  • Applications: Western blot, immunohistochemistry, and ELISA*
    • *The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

HRP Datasheet

AP (Alkaline Phosphatase)

Properties:

  • Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenols
  • Commonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligation
  • Catalyzes the conversion of:
    • Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured products
    • Fluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent products
  • Molecular weight: 140 kDa
  • Applications: Western blot, immunohistochemistry, and ELISA

AP Datasheet

  R-PE (R-Phycoerythrin)
Overview:

  • Broad excitation spectrum
  • High quantum yield
  • Photostable
  • Member of the phycobiliprotein family
  • Isolated from red algae
  • Excellent solubility in water
  • Molecular Weight: 250 kDa

 

  • Applications: The listed applications provide a general overview of potential uses for conjugated antibodies. However, they do not guarantee that every antibody-conjugate combination has been tested or validated for these specific applications.

R-PE Datasheet

  R-PE Fluorophore Excitation and Emission Spectra Optical Properties:

λex = 565 nm

λem = 575 nm

εmax = 2.0×106

Φf = 0.84

Brightness = 1.68 x 103

Laser = 488 to 561 nm

Filter set = TRITC

 
Field of Use Not for use in humans. Not for use in diagnostics or therapeutics. For in vitro research use only.

Properties

Storage Buffer PBS pH 7.4, 50% glycerol, 0.1% sodium azide *Storage buffer may change when conjugated
Storage Temperature -20ºC, Conjugated antibodies should be stored according to the product label
Shipping Temperature Blue Ice or 4ºC
Purification Protein G Purified
Clonality Monoclonal
Clone Number 4B4
Isotype IgG1
Specificity Detects ~85kDa (unstressed cell lysates) and ~95kDa (heat shocked cell lysates).
Cite This Product StressMarq Biosciences Cat# SMC-477, RRID: AB_2702297
Certificate of Analysis 1 µg/ml of SMC-477 was sufficient for detection of HSF1 in 20 µg of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Rabbit anti-rat IgG: AP as the secondary antibody.

Biological Description

Alternative Names HSTF1 Antibody, Heat shock factor protein 1 Antibody, Heat shock transcription factor 1 Antibody, HSF 1 Antibody
Research Areas Cancer, Cardiovascular System, Cell Signaling, Epigenetics and Nuclear Signaling, Heart, Heat Shock
Cellular Localization Cytoplasm, Nucleus
Accession Number NP_032322.1
Gene ID 15499
Swiss Prot P38532
Scientific Background HSF1, or heat shock factor 1, belongs to a family of Heat Shock transcription factors that activate the transcription of genes encoding products required for protein folding, processing, targeting, degradation, and function (2). The up-regulation of HSP (heat shock proteins) expression by stressors is achieved at the level of transcription through a heat shock element (HSE) and a transcription factor (HSF) (3, 4, 5). Most HSFs have highly conserved amino acid sequences. On all HSFs there is a DNA binding domain at the N-terminus. Hydrophobic repeats located adjacent to this binding domain are essential for the formation of active trimers. Towards the C-terminal region another short hydrophobic repeat exists, and is thought to be necessary for suppression of trimerization (6). There are two main heat shock factors, 1 and 2. Mouse HSF1 exists as two isoforms, however in higher eukaryotes HSF1 is found in a diffuse cytoplasmic and nuclear distribution in un-stressed cells. Once exposed to a multitude of stressors, it localizes to discrete nuclear granules within seconds. As it recovers from stress, HSF1 dissipates from these granules to a diffuse nuceloplasmic distribution. HSF2 on the other hand is similar to mouse HSF1, as it exists as two isoforms, the alpha form being more transcriptionally active than the smaller beta form (7, 8). Various experiments have suggested that HFS2 may have roles in differentiation and development (9, 10, 11).
References 1. Cotto J.J., Fox S.G. and Morimoto R.I. (1997) J. Cell Science 110: 2925-2934.
2. Morano K.A. and Thiele D.J. (1999). Gene Expression 7 (6): 271-82.
3. Tanaka K.I., et al. (2007). JBC Papers Online Manuscript M704081200.
4. Morimoto R. I. (1998) Genes Dev 12: 3788-3796.
5. McMillan D. R., et al. (1998) J Bio Chem 273: 7523-7528.
6. Jolly C., Usson Y. and Morimoto R.I. (1999) Proc. Natl. Acad. Sci. USA 96 (12): 6769- 6774.
7. Fiorenza M.T., et al. (1995) Nucleic Acids Res. 23 (3):467-474.
8. Goodson M.L., Park-Sarge O.K. and Sarge K.D. (1995) Mol. Cell. Biol. 15(10): 5288-5293.
9. Rallu M., et al. (1997) Proc. Natl. Acad. Sci. USA 94(6): 2392-2397.
10. Sarge K.D., et al. (1994) Biol. Reprod. 50(6): 1334-1343.
11. Murphy S.P., Gorzowski J.J., Sarge K.D. and Phillips B. (1994) Mol. Cell. Biol. 14(8):5309-5317.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Heat Shocked cervical cancer cells (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rat (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Localizes to the nucleus upon activation. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-HSF1 Antibody. (C) Composite. Heat Shocked at 42°C for 1h.</p>

Immunocytochemistry/Immunofluorescence analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Heat Shocked cervical cancer cells (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rat (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Localizes to the nucleus upon activation. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-HSF1 Antibody. (C) Composite. Heat Shocked at 42°C for 1h.

<p>Immunocytochemistry/Immunofluorescence analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Heat Shocked cervical cancer cells (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:100 for 12 hours at 4°C. Secondary Antibody: APC Goat Anti-Rat (red) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Localizes to the nucleus upon activation. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-HSF1 Antibody. (C) Composite. Heat Shocked at 42°C for 1h.</p>

Immunocytochemistry/Immunofluorescence analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Heat Shocked cervical cancer cells (HeLa). Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:100 for 12 hours at 4°C. Secondary Antibody: APC Goat Anti-Rat (red) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Localizes to the nucleus upon activation. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-HSF1 Antibody. (C) Composite. Heat Shocked at 42°C for 1h.

<p>Immunohistochemistry analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Breast carcinoma. Species: Human. Fixation: 10% Formalin Solution for 20 hours at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:2000 for 40 min. Secondary Antibody: Dako labeled Polymer HRP Anti-rat IgG, DAB Chromogen (brown) (Dako Envision+ System) for 30 min at RT. Counterstain: Mayer’s Hematoxylin (purple/blue) nuclear stain for 1 minute at RT. Localization: Nuclear. Magnification: 100X. Courtesy of: Dr. Sandro Santagata, Harvard Medical School.</p>

Immunohistochemistry analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Breast carcinoma. Species: Human. Fixation: 10% Formalin Solution for 20 hours at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:2000 for 40 min. Secondary Antibody: Dako labeled Polymer HRP Anti-rat IgG, DAB Chromogen (brown) (Dako Envision+ System) for 30 min at RT. Counterstain: Mayer’s Hematoxylin (purple/blue) nuclear stain for 1 minute at RT. Localization: Nuclear. Magnification: 100X. Courtesy of: Dr. Sandro Santagata, Harvard Medical School.

<p>Western Blot analysis of Human Breast adenocarcinoma cell line (MCF7) showing detection of ~65 kDa HSF1 protein using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Lane 1: MW ladder. Lane 2: HSF1 null lysate prepared from mouse embryonic fibroblasts. Lane 3: MCF7 lysate (5 µg). Lane 4: MCF7 lysate (10 µg). Lane 5: MCF7 lysate (20 µg). Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Rat IgG: HRP for 1 hour at RT. Predicted/Observed Size: ~65 kDa. Courtesy of: Dr. Sandro Santagata, Harvard Medical School.</p>

Western Blot analysis of Human Breast adenocarcinoma cell line (MCF7) showing detection of ~65 kDa HSF1 protein using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Lane 1: MW ladder. Lane 2: HSF1 null lysate prepared from mouse embryonic fibroblasts. Lane 3: MCF7 lysate (5 µg). Lane 4: MCF7 lysate (10 µg). Lane 5: MCF7 lysate (20 µg). Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Rat IgG: HRP for 1 hour at RT. Predicted/Observed Size: ~65 kDa. Courtesy of: Dr. Sandro Santagata, Harvard Medical School.

<p>Immunohistochemistry analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Lung. Species: Mouse. Fixation: 10% Formalin Solution for 20 hours at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:1000 for 40 min. Secondary Antibody: Dako labeled Polymer HRP Anti-rat IgG, DAB Chromogen (brown) (Dako Envision+ System) for 30 min at RT. Counterstain: Mayer’s Hematoxylin (purple/blue) nuclear stain for 1 minute at RT. Localization: Nuclear. Magnification: 100X. (A) HSF Wildtype. (B) HSF null. Courtesy of: Dr. Sandro Santagata, Harvard Medical School.</p>

Immunohistochemistry analysis using Rat Anti-HSF1 Monoclonal Antibody, Clone 4B4 (SMC-477). Tissue: Lung. Species: Mouse. Fixation: 10% Formalin Solution for 20 hours at RT. Primary Antibody: Rat Anti-HSF1 Monoclonal Antibody (SMC-477) at 1:1000 for 40 min. Secondary Antibody: Dako labeled Polymer HRP Anti-rat IgG, DAB Chromogen (brown) (Dako Envision+ System) for 30 min at RT. Counterstain: Mayer’s Hematoxylin (purple/blue) nuclear stain for 1 minute at RT. Localization: Nuclear. Magnification: 100X. (A) HSF Wildtype. (B) HSF null. Courtesy of: Dr. Sandro Santagata, Harvard Medical School.

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