Overview

  • Product name
    Anti-Hsp70 antibody [EP1007Y]
    See all Hsp70 primary antibodies
  • Description
    Rabbit monoclonal [EP1007Y] to Hsp70
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide within Human Hsp70 aa 600-700 (C terminal). The exact sequence is proprietary.

  • Positive control
    • HeLa whole cell lysate (ab150035),human prostate carcinoma.
  • General notes

    A trial size is available to purchase for this antibody.

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab45133 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF 1/400 - 1/1000.
WB 1/1000 - 1/10000. Detects a band of approximately 85 kDa.
IHC-P 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Relevance
    Function: In cooperation with other chaperones, the Hsp70 family stabilize preexistent proteins against aggregation and mediate the folding of newly translated polypeptides in the cytosol as well as within organelles. These chaperones participate in all these processes through their ability to recognize nonnative conformations of other proteins. They bind extended peptide segments with a net hydrophobic character exposed by polypeptides during translation and membrane translocation, or following stress-induced damage. In case of rotavirus A infection, serves as a post-attachment receptor for the virus to facilitate entry into the cell. Tissue specificity: HSPA1B is testis-specific.
  • Cellular localization
    Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Database links
  • Alternative names
    • DnaK type molecular chaperone HSP70 1 antibody
    • Epididymis secretory protein Li 103 antibody
    • FLJ54303 antibody
    • FLJ54370 antibody
    • FLJ54392 antibody
    • FLJ54408 antibody
    • FLJ75127 antibody
    • Heat shock 70 kDa protein 1 antibody
    • Heat shock 70 kDa protein 1/2 antibody
    • Heat shock 70 kDa protein 1A antibody
    • Heat shock 70 kDa protein 1A/1B antibody
    • Heat shock 70kDa protein 1B antibody
    • Heat shock induced protein antibody
    • HEL S 103 antibody
    • HSP70 1 antibody
    • HSP70 1A antibody
    • HSP70 1B antibody
    • HSP70 2 antibody
    • HSP70-1 antibody
    • HSP70-1/HSP70-2 antibody
    • HSP70-1A antibody
    • HSP70.1 antibody
    • HSP70.1/HSP70.2 antibody
    • HSP71_HUMAN antibody
    • HSP72 antibody
    • HSPA1 antibody
    • HSPA1A antibody
    • HSPA1B antibody
    see all

Images

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labelling Hsp70 with purified ab45133 at 1/1000. Cells were fixed with 100% methanol and permeabilized with 0.1% triton X-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.

  • Immunohistochemical staining of paraffin embedded human breast carcinoma with purified ab45133 at a working dilution of 1 in 400. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • All lanes : Anti-Hsp70 antibody [EP1007Y] (ab45133) at 1/10000 dilution (purified)

    Lane 1 : A431 cell lysate
    Lane 2 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Observed band size: 70 kDa
    why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Overlay histogram showing Jurkat cells stained with unpurified ab45133 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab45133, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • Unpurified ab45133.

References

This product has been referenced in:
  • Borroni AP  et al. Smurf2 regulates stability and the autophagic-lysosomal turnover of lamin A and its disease-associated form progerin. Aging Cell 17:N/A (2018). Read more (PubMed: 29405587) »
  • Yu B  et al. Diagnostic potential of serum exosomal colorectal neoplasia differentially expressed long non-coding RNA (CRNDE-p) and microRNA-217 expression in colorectal carcinoma. Oncotarget 8:83745-83753 (2017). Read more (PubMed: 29137379) »
See all 12 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Question
Answer

Vielen Dank, dass Sie uns kontaktiert haben und für Ihre Geduld, während ich das Labor um Hilfe anfragte.
Das Labor konnte leider auch nicht erklären, woher Ihre grosse Bande kommt. In der Tat ist in der Litterature die Bande auch bei Cos7 Zellen eher normal. Siehe folgende Publikation: http://jmg.bmj.com/content/41/1/47.full
Das Labor hat beim testen ähnlicher Zellen, den COS-1 Zellen, auch nur eine Bande entdeckt - siehe Anhang. Wieso Sie in den Cos7 Zellen eine solche Bande sehen, ist mir deshalb nicht klar.
Eine Hypothese wäre, dass die COS7 Zellen das Hsp70 verstärkt glycolysieren. Falls Sie dies testen möchten, könnten Sie das Zelllysate mit den entsprechenden Glycosidasen behandeln. Es gibt auch kommerzielle Kits die dies machen.
Das Labor hat die exakte Sequenz mit dem HSPA2 SwissProt P54652 aligniert. Die Identität beträgt 80%. Die Wahrscheinlichkeit einer Kreuzreaktion ist daher eher gering, kann jedoch nicht ganz ausgeschlossen werden.
Um eine limitierte Proteolyse auch auszuschliessen, kann ich auch empfehlen einen Proteaseinhibitorencocktail dem Lysat hinzuzufügen und das Lysat immer auf Eis behalten.
Leider haben wir kein Cos7 Kontrolllysat in unserem Katalog welches wir Ihnen senden könnten. Wir haben nur das nukleare Lysat.
Haben Sie unterdessen weitere Daten mit diesem Antikörper bekommen, welche diese Banden besser erklären könnten?
Ich freue mich Ihre Kommentare zu den obigen Punkten zu hören. Bitte lassen Sie mich auch wissen, falls Sie irgendwelche anderen Fragen oder Bedenken haben.
Ich freue mich wieder von Ihnen zu hören.

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Question
Answer

Vielen Dank für Ihren Anruf.

Es tut mir leid zu hören, dass Sie Probleme mit diesem Antikörper haben.

Ich habe unseren Fragebogen als Word-Dokument an diese E-Mail angehängt. Durch das Ausfüllen des Fragebogens erhalten wir alle nötigen Informationen über Ihre Proben und Ihr Protokoll. Sobald Sie dieses Formular an uns zurückgeschickt haben, werden wir uns Ihr Protokoll ansehen und versuchen herauszufinden, wieso die Bande die Sie sehen so breit ist. Falls sich herausstellt, dass der Antikörper nicht so funktioniert, wie auf dem Datenblatt beschrieben und er innerhalb der letzten 180 Tage gekauft wurde, werden wir Ihnen gerne einen Ersatz oder eine Gutschrift schicken.

Ich freue mich, bald wieder von Ihnen zu hören.

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Question
Answer

Jederzeit gerne wieder :-)

Ihnen auch ein schönes Wochenende!

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