Anti-HSP70 Antibody [5A5]

Mouse Anti-Human HSP70 Monoclonal IgG1

Catalog No. SMC-162

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Reviews (1) | Datasheet
Species Reactivity Hu, Ms, Rt, Am, Ck, Fs, Ys, Dr
Applications WB IHC ICC/IF FCM IP
SKU: SMC-162 Categories: ,

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SMC-162-HSP70-Antibody-5A5-ICC-IF-Human-Cervical-Cancer-cell-line-HeLa-40X-Composite-1.png
Mouse Anti-Hsp70 Antibody [5A5] used in Western Blot (WB) on Rat skeletal muscle tissue lysate (SMC-162)
Product Name HSP70 Antibody
Description

Mouse Anti-Human HSP70 Monoclonal IgG1
Species Reactivity Amphibians, Chicken, Fish, Fruit Fly (Drosophila melanogaster), Human, Mouse, Rat, Yeast, Yeast (Saccharomyces cerevisiae)
Applications WB, IHC, ICC/IF, IP, AM
Antibody Dilution WB (1:1000), ICC/IF (1:500), IP (1µg); optimal dilutions for assays should be determined by the user.
Host Species Mouse
Immunogen Species Human
Immunogen Human recombinant HSP70 overexpressed in E.coli
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

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

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

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 

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 

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

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

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

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

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 pH7.2, 50% glycerol, 0.09% 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 5A5
Isotype IgG1
Specificity Detects ~70kDa. May detect HSP70, HSC70, Grp78 and HSP72.
Cite This Product StressMarq Biosciences Cat# SMC-162, RRID: AB_2119819
Certificate of Analysis 1 µg/ml of SMC-162 was sufficient for detection of HSP70 in 20 µg of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody.

Biological Description

Alternative Names HSP70 1 Antibody, HSP70 2 Antibody, HSP70.1 Antibody, HSP72 Antibody, HSPA1 Antibody, HSPA1A Antibody, HSPA1B Antibody
Research Areas Cancer, Cell Signaling, Chaperone Proteins, Heat Shock, Protein Trafficking, Tumor Biomarkers
Cellular Localization Cytoplasm
Accession Number NP_005336.3
Gene ID 3303
Swiss Prot P0DMV8/P0DMV9
Scientific Background HSP70 genes encode abundant heat-inducible 70-kDa HSPs (HSP70s). In most eukaryotes HSP70 genes exist as part of a multigene family. They are found in most cellular compartments of eukaryotes including nuclei, mitochondria, chloroplasts, the endoplasmic reticulum and the cytosol, as well as in bacteria. The genes show a high degree of conservation, having at least 50% identity (2). The N-terminal two thirds of HSP70s are more conserved than the C-terminal third. HSP70 binds ATP with high affinity and possesses a weak ATPase activity which can be stimulated by binding to unfolded proteins and synthetic peptides (3). When HSC70 (constitutively expressed) present in mammalian cells was truncated, ATP binding activity was found to reside in an N-terminal fragment of 44 kDa which lacked peptide binding capacity. Polypeptide binding ability therefore resided within the C-terminal half (4). The structure of this ATP binding domain displays multiple features of nucleotide binding proteins (5). All HSP70s, regardless of location, bind proteins, particularly unfolded ones. The molecular chaperones of the HSP70 family recognize and bind to nascent polypeptide chains as well as partially folded intermediates of proteins preventing their aggregation and misfolding. The binding of ATP triggers a critical conformational change leading to the release of the bound substrate protein (6). The universal ability of HSP70s to undergo cycles of binding to and release from hydrophobic stretches of partially unfolded proteins determines their role in a great variety of vital intracellular functions such as protein synthesis, protein folding and oligomerization and protein transport. For more information visit our HSP70 Scientific Resource Guide at http://www.HSP70.com.
References 1. Balashova N. et al. (2005) J Biol Chem 280:2186-96.
2. Boorstein W. R., Ziegelhoffer T. & Craig E. A. (1993) J. Mol. Evol.38 (1): 1-17.
3. Rothman J. (1989) Cell 59: 591 -601.
4. DeLuca-Flaherty et al. (1990) Cell 62: 875-887.
5. Bork P., Sander C. & Valencia A. (1992) Proc. Nat Acad. Sci. USA 89: 7290-7294.
6. Fink A.L. (1999) Physiol. Rev. 79: 425-449.

Product Images

<p>Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 5A5 (SMC-162). Tissue: Cervical cancer cell line (HeLa). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-162) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Nucleus, Cytoplasm. Magnification: 40X.</p>

Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 5A5 (SMC-162). Tissue: Cervical cancer cell line (HeLa). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-162) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Nucleus, Cytoplasm. Magnification: 40X.

<p>Western Blot analysis of Rat skeletal muscle tissue lysate showing detection of Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone 5A5 (SMC-162). Load: 15 µg. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody (SMC-162) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.</p>

Western Blot analysis of Rat skeletal muscle tissue lysate showing detection of Hsp70 protein using Mouse Anti-Hsp70 Monoclonal Antibody, Clone 5A5 (SMC-162). Load: 15 µg. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp70 Monoclonal Antibody (SMC-162) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT.

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