Anti-HSP70 Antibody [1H11]

Mouse Anti-Human Cell Membrane HSP70 Monoclonal IgG1

Catalog No. SMC-249

5 out of 5 based on 1 customer rating
Species Reactivity , Hu , Ms , Rt , Bv , Pg
Applications WB IHC ICC/IF FCM IP
5 out of 5 based on 1 customer rating
(1 customer review)

USD $40.00USD $404.00

SKU: SMC-249 Categories: , .

Next day delivery if ordered before 12pm PST

Clear
SMC-249-HSP70-Antibody-1H11-ICC-IF-Human-HCT116-cells-1.png
Mouse Anti-HSP70 Antibody: FITC [1H11] used in Fluorescence-activated cell sorting (FACS) on Human Jurkat E6.1 cells (SMC-249)Mouse Anti-HSP70 Antibody [1H11] used in Western Blot (WB) on Human Heat shocked HeLa cell lysates (SMC-249)Mouse Anti-HSP70 Antibody: FITC [1H11] used in Immunocytochemistry/Immunofluorescence (ICC/IF) on Human HCT116 cells (SMC-249)Mouse Anti-HSP70 Antibody: FITC [1H11] used in Fluorescence-activated cell sorting (FACS) on Human HCT116 and HK2 cells (SMC-249)Mouse Anti-HSP70 Antibody: FITC [1H11] used in Fluorescence-activated cell sorting (FACS) on Human Jurkat E6.1, U937, MCF7 and HT29 cells (SMC-249)

Overview

Product Name HSP70 Antibody
Description

Mouse Anti-Human Cell Membrane HSP70 Monoclonal IgG1

Species Reactivity Human, Mouse, Rat, Bovine, Pig
Applications , WB , ICC/IF , FCM , FACS
Antibody Dilution WB (1:1000), ICC/IF (1:100), FACS (1:250); optimal dilutions for assays should be determined by the user.
Host Species Mouse
Immunogen Species Human
Immunogen Human native HSP70 protein
Concentration 1 mg/ml
Conjugates Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated
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

 

  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

 

  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 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 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 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 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 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 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

 

  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

 

 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

 

 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

 

  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

 

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

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

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

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

Properties

Storage Buffer PBS pH 7.4, 50% glycerol, 0.9% Sodium Azide
Storage Temperature -20ºC
Shipping Temperature Blue Ice or 4ºC
Purification Protein G Purified
Clonality Monoclonal
Clone Number 1H11
Isotype IgG1
Specificity Binds to cell surface HSP70 without permeabilization of cell membrane in tumor cell lines. Does not bind to intracellular HSP70 at low concentrations. Detects a ~70 kDa band in Western Blot.
Cite This Product Mouse Anti-Human HSP70 Monoclonal (StressMarq Biosciences Inc., Victoria BC CANADA, Catalog # SMC-249)
Certificate of Analysis A 1:250 dilution of SMC-249 was sufficient for detection of cell surface HSP70 in HCT116 cells using Fluorescence-activated cell sorting, applied in flow cytometry, with FITC as the fluorescent probe.

Biological Description

Alternative Names HSP70 1 Antibody, HSP70 2 Antibody, HSP70.1 Antibody, HSP72 Antibody, HSPA1 Antibody, HSPA1A Antibody, HSPA1B Antibody
Research Areas Cancer, Heat Shock, Cell Signaling, Chaperone Proteins, Protein Trafficking, Tumor Biomarkers
Cellular Localization Cell membrane
Accession Number NP_005336.3
Gene ID 3303
Swiss Prot P0DMV8
Scientific Background HSP70 is a highly conserved protein that is ubiquitously expressed. It can be found in chloroplasts, eukaryotic cytosol, endoplasmic reticulum, and mitochondria, but also embedded in the cell membrane and in the extracellular space (1,2,3,4). Even though HSP70 is one of the most studied heat shock proteins, the export mechanism and method of membrane insertion are not fully understood. The majority of the proteins in the heat shock family lack a consensus signal for secretion via the ER-Golgi pathway (5). Researchers have found that HSP70 may be released from cells via exosomes or secretory vesicles (6).
Although HSP70 is ubiquitously expressed, there is not much information about its presence on cell surface. The finding that HSP70 is localized on the cell surface of cancer cells but not normal cells suggests a conformational change of HSP70 in the lower pH environment characteristic of cancer cells (7). The presence of cell membrane embedded HSP70 has been found to increase the stability of cancer cells, thereby protecting tumors from environmental stress (8, 9).
Anti-HSP70 antibody, clone 1H11 (Catalog No. SMC-249) is unique from other commercially available antibodies in that it can bind to the extracellular region of the cell membrane embedded HSP70 protein, allowing researchers to differentiate between membrane bound and intracellular HSP70 across cancer cells types.
References 1. Gribaldo, S. et al. (1999) J. Bacteriol. 181, 434-443.
2. Sharma, D. & Masison, D.C. (2009) Protein Pept. Lett. 16, 571-581.
3. Sharma, D. et al. (2009) PLoS. ONE.4, e6644.
4. Kampinga, H.H. & Craig, E.A. (2010) Nat. Rev. Mol. Cell Biol. 11, 579-592.
5. Nickel, W. & Seedorf, M. (2008) Annu. Rev. Cell Dev. Biol. 24, 287-308.
6. Multhoff, G. & Hightower, L.E. (1996) Cell Stress. Chaperones. 1, 167-176.
7. De Maio, A. (2011) Cell Stress. Chaperones. 16, 235-249.
8. Horvath, I. & Vigh, L. (2010) Nature. 463, 436-438.
9. Horvath, I., Multhoff, G., Sonnleitner, A., & Vigh, L. (2008) Biochim. Biophys. Acta 1778, 1653-1664.

Product Images

Immunocytochemistry/Immunofluorescence analysis on non-permeabilized HCT116 cells using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC) showing cell membrane staining. Tissue: HCT116 cells. Species: Human. Fixation: 4% Formaldehyde. Permeabilization: None. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100. Counterstain: Wheat germ agglutinin Texas red membrane marker; DAPI (blue) nuclear stain. Localization: Cell surface, cell membrane. (A) DAPI nuclear stain. (B) Wheat germ agglutinin Texas red. (C) HSP70 Antibody. (D) Composite. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut.

Fluorescence-activated cell sorting (FACS) using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: Jurkat E6.1. Species: Human. Unfixed/non-permeablized cells were incubated with (A) Anti-HSP70 control antibody, or (B) Anti-HSP70 antibody, clone 1H11 (SMC-249D-FITC), for 40 min at 4°C. Fixed/permeabilized cells were incubated with (C) Anti-HSP70 control antibody, or (D) Anti-HSP70 antibody, clone 1H11 (SMC-249D-FITC) for 40 min at 4°C. Analysis shows that Anti-HSP70 (SMC-249) can detect low levels of cell surface HSP70, and can fainlty detect intracellular HSP70. Courtesy of: Dr. Elyse Ireland, Institute of Medicine, University of Chester.

Western Blot analysis of Human Heat shocked HeLa cell lysates showing detection of HSP70 protein using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11 (SMC-249). Lane 1: Molecular Weight ladder (MW). Lane 2: HeLa cell lysates. Load: 20 µg. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:1000.

Immunocytochemistry/Immunofluorescence analysis on permeabilized HCT116 cells using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC) showing faint cell membrane and intracellular staining. Tissue: HCT116 cells. Species: Human. Fixation: 4% Formaldehyde. Permeabilization: 0.2% PBST. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100. Counterstain: Wheat germ agglutinin Texas red membrane marker; DAPI (blue) nuclear stain. Localization: Cell surface, faint intracellular. (A) DAPI nuclear stain. (B) Wheat germ agglutinin Texas red. (C) HSP70 Antibody. (D) Composite. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut.

Fluorescence-activated cell sorting analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: HCT116 and HK2 cells. Species: Human. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 1:100 for 90 min at 4°C. Counterstain: Propidium Iodide nuclear stain. (A) HK2 cells unstained. (B) HK2 cells and HSP70 Antibody. (C) HCT116 cells unstained. (D) HCT116 cells and HSP70 Antibody. Shows that HSP70 Antibody binds to the cell surface of tumor cells but not non-tumor cells. Courtesy of: Lawrence Hightower, Charles Giardina, and Didem Ozcan from University of Connecticut.

Fluorescence-activated cell sorting analysis using Mouse Anti-HSP70 Monoclonal Antibody, Clone 1H11: FITC conjugate (SMC-249D-FITC). Tissue: Jurkat E6.1, U937, MCF7 and HT29 cells . Species: Human. Fixation: No fixation. Permeabilization: None. Primary Antibody: Mouse Anti-HSP70 Monoclonal Antibody (SMC-249) at 20 µg/ml for 40 min at 4°C. Counterstain: Propidium Iodide nuclear stain at 2.5 µg/ml for 5 min at 4°C. Shows that binding of HSP70 Antibody is not cell-line specific. Courtesy of: Dr. Elyse Ireland, Institute of Medicine, University of Chester.

Product Citations (1)

Other Citations

Extracellular Hsp70 Enhances Mesoangioblast Migration via an Autocrine Signaling Pathway.

Barreca, M.M. et al. (2016) J Cell Physiol. [Epub ahead of print].

PubMed ID: 27925208 Reactivity: Mouse Applications: Protein Inhibition

Reviews

1 review for HSP70 Antibody

  1. 5 out of 5

    :

    Based on validation through cited publications.

Add a review