Anti-Hsp70 antibody [N27F3-4] (ab47454)

Overview

  • Product nameAnti-Hsp70 antibody [N27F3-4]See all Hsp70 primary antibodies ...
  • Description
    Mouse monoclonal [N27F3-4] to Hsp70
  • SpecificityDetects 72 and 73kDa proteins corresponding to the molecular mass of inducible Hsp and Hsc70 of SDS PAGE immunoblots.
  • Tested applicationsIHC-Fr, IHC-P, WB, IP, Flow Cyt, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Dog, Human, Pig, Xenopus laevis, Caenorhabditis elegans, Carp, Fruit fly (Drosophila melanogaster), Fish, Monkey
    Predicted to work with: Plants
  • Immunogen

    Recombinant hsp70/hsc70 (Human)

  • Positive control
    • HeLa lysate IF/ICC: Hek293 celll ine

Properties

Applications

Our Abpromise guarantee covers the use of ab47454 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 70 kDa.
IP Use a concentration of 5 µg/ml.
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

Target

Anti-Hsp70 antibody [N27F3-4] images

  • Hsp70 was immunoprecipitated using 0.5mg Rat Testis tissue lysate, 5µg of Mouse monoclonal to Hsp70 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10min, Rat Testis tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.

    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab47454.

    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/20,000 dilution.

    Band: 70kDa, non specific bands - 50kDa: We are unsure as to the identity of this extra band; Hsp70

  • Anti-Hsp70 antibody [N27F3-4] (ab47454) at 1/500 dilution + Apteronotus leptorhynchus brain tissue lysate at 50 µg

    Secondary
    Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal at 1/1000 dilution

    Performed under reducing conditions.

    Predicted band size : 70 kDa
    Observed band size : 70 kDa


    Exposure time : 5 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-Hsp70 antibody [N27F3-4] (ab47454) at 1 µg/ml

    Lane 1 : Cell lysate prepared from human A431 cell line
    Lane 2 : Cell lysate prepared from human A549 cell line
    Lane 3 : Cell lysate prepared from human HCT116 cell line
    Lane 4 : Cell lysate prepared from human Hela cell line
    Lane 5 : Cell lysate prepared from human HEK293 cell line
    Lane 6 : Cell lysate prepared from human HepG2 cell line
    Lane 7 : Cell lysate prepared from human HL-60 cell line
    Lane 8 : Cell lysate prepared from human HUVEC cell line
    Lane 9 : Cell lysate prepared from human Jurkat cell line
    Lane 10 : Cell lysate prepared from human MCF7 cell line
    Lane 11 : Cell lysate prepared from human PC3 cell line
    Lane 12 : Cell lysate prepared from human T98G cell line
    Lane 13 : Lysate prepared from rat brain tissue

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 70 kDa
  • ICC/IF image of ab47454 stained Hek293 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab47454, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Hsp70 antibody [N27F3-4] (ab47454)

ab47454 has not yet been referenced specifically in any publications.

Product Wall

Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Antigen retrieval step None
Sample Apteronotus leptorhynchus Tissue sections (Brain)
Specification Brain
Permeabilization Yes - 0.3% Triton X-100
Fixative Formaldehyde
Username

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Submitted Aug 29 2013

Application Western blot
Loading amount 50 µg
Gel Running Conditions Reduced Denaturing (12%)
Sample Apteronotus leptorhynchus Tissue lysate - whole (Brain)
Specification Brain
Blocking step Milk as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Submitted Aug 29 2013

Thank you for your enquiries.

I am now back in the office and have reviewed this case for you. Regrettably, we have no other Hsp70 antibodies that have been tested in Drosophila or that have specifically been tested andguaranteed not to reac...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (HeLa and U2OS Cells)
Loading amount 30 µg
Specification HeLa and U2OS Cells
Gel Running Conditions Reduced Denaturing (12% Tris Glycine)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

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Verified customer

Submitted Jan 18 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"