Overview

  • Product name
    Anti-Hsp70 antibody [C92F3A-5]
    See all Hsp70 primary antibodies
  • Description
    Mouse monoclonal [C92F3A-5] to Hsp70
  • Host species
    Mouse
  • Specificity
    Detects a 70kDa protein corresponding to the molecular mass of Hsp70 of SDS PAGE immunoblots. There is no cross-reactivity to Hsc70 (Hsp73).
  • Tested applications
    Suitable for: Electron Microscopy, WB, IP, ELISA, ICC/IF, Flow Cyt, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Rat, Human
    Predicted to work with: Mouse, Cow, Dog, Pig, Caenorhabditis elegans, Macaque monkey, Bos mutus grunniens, Saguinus oedipus, Onchocerca volvulus, Oncorhynchus tshawytscha
  • Immunogen

    Recombinant fragment corresponding to Human Hsp70 aa 436-503.
    Sequence:

    YSDNQPGVLIQVYEGERAMTKDNNLLGRFELSGIPPAPRGVPQIEVTFDI DANGILNVTATDKSTGKANKITI


    Database link: P08107

  • Epitope
    The mapped epitope is in the region of amino acid residues 436-503.
  • Positive control
    • WB: HeLa, A431, A549, HCT116, HEK293, HepG2, HL-60, HUVEC, Jurkat, MCF7, PC3, T98G cell lysates; Rat Brain tumour lysate; Rat bone lysate. IHC-P: Mouse colon cancer tissue; human colon cancer tissue. Flow cyt: HeLa cells.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.20
    Preservative: 0.09% Sodium azide
    Constituents: 50% PBS, 50% Glycerol
  • Concentration information loading...
  • Purity
    Protein G purified
  • Clonality
    Monoclonal
  • Clone number
    C92F3A-5
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab47455 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
Electron Microscopy Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 70 kDa.
IP Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

IHC-Fr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.

Target

Images

  • All lanes : Anti-Hsp70 antibody [C92F3A-5] (ab47455) 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 cells 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 : Rat brain tissue lysates

    Predicted band size: 70 kDa



    All are cancer cell lines.

  • Western blot of Hela cell heat shock lysate, using HRP conjugated goat anti-mouse secondary antibody for detection. (1µg/mL ab47455 was used to detect in 10µg of heat shocked HeLa cell lysate)

  • Formalin-fixed, paraffin-embedded human colon carcinoma tissue stained for Hsp70 using ab47455 at 1/1000 dilution in immunohistochemical analysis. Counter stained with hematoxylin.

  • All lanes : Anti-Hsp70 antibody [C92F3A-5] (ab47455) at 1/500 dilution

    All lanes : Rat bone (tibia) whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Predicted band size: 70 kDa

    See Abreview

  • ab47455 staining Hsp70 in mouse colon cancer tissue section by Immunohistochemistry (Bouin's fixed paraffin embedded tissue sections). The primary antibody was diluted at 1/100,000. A Fluorophore conjugated goat anti mouse was used as secondary.  An antibody amplifier™ system was used for staining.

  • Overlay histogram showing HeLa cells stained with ab47455 (red line). The cells were fixed with methanol (5 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 (ab47455, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1:500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a significantly decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:
  • Zhu X  et al. Upconversion nanocomposite for programming combination cancer therapy by precise control of microscopic temperature. Nat Commun 9:2176 (2018). Read more (PubMed: 29872036) »
  • Benitez-Amaro A  et al. Identification of new biophysical markers for pathological ventricular remodelling in tachycardia-induced dilated cardiomyopathy. J Cell Mol Med 22:4197-4208 (2018). Read more (PubMed: 29921039) »
See all 21 Publications for this product

Customer reviews and Q&As

1-10 of 17 Abreviews or Q&A

Application
Western blot
Sample
Rat Tissue lysate - whole (Bone tissue (Tibia))
Gel Running Conditions
Reduced Denaturing (12,5%)
Loading amount
50 µg
Specification
Bone tissue (Tibia)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Aug 25 2015

Question

thank you very much for your reply.
One last question, in totally normal cells will I get a band of HSP 70 (not HSP73) in WB? I learned that HSP 73 is constitutionally produced by cells, while HSP70 protein is induced by heat, injury, which makes me to think that HSP70 would not appear in normal cell. Is this correct?














https://www.abcam.com/index.html?utm_campaign=CRM&utm_source=Abcam.CRM&utm_medium=Email&utm_term=Header








Dear Sir/Madam








Thank you for contacting us.

Yes, this product has been tested sucessfully in rabbit for Western Blotting. Our results show a clean band.

The antibody should specifically detect HSP70 and not HSP73.

Regarding local injury, I am not sure whether your levels will be elevated or not. Cells that are heat shocked do increase HSP70 so it seems reasonable that this would appear in your staining but this would have to be tested.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.



Help us improve our service.
https://www.abcam.com/index.html?pageConfig=technicalSurvey&intCCEID=4637456





Best regards,
Heather

Heather Allen
Scientific Support Specialist
Abcam Inc.
https://www.abcam.com










Your original inquiry to Abcam:


Sorry to bother you, but really need help.



I bought the HSP70 antibody from your company. The product is Anti-Hsp70 antibody [C92F3A-5] ab47455.

I have a few questions about the functions of the antibody. Some of the answers to them may seem easy to find on internet, but my english is not good enough, finding answers will be very annoying to me. So I want to ask for help from you. Thank you very much!



1, Is it applicable in rabbit experiment? I read the specification of the product, but I am still not sure about the question. I mean when I am doing a rabbit experiment, and I want to detect the HSP70 (induced by pressure), not HSP73 (constitutionally existing in cells), is it correct to use this product?

2,Does the HSP70 ( not HSP73) present in normal cells. I mean if I measure the quantity of the HSP70 protein in a totally normal cell, will I get the protein band in Western blot? Will the band be very blurring? Or clear?

3,If I add a traumatic stimulant to the normal rabbit brain, I want to detect the HSP70 protein quantity around the lesion. Will I get the elevated HSP70 result? I read some papers on HSP70, and the elevated HSP70 results were all obtained by whole body overheated not just local injury. So I doubt the effect of the local injury.

Thank you very much for your patience.






[CCE4637456]

SID:42













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Answer

In normal cells there is still some low expression of HSP70, which is upregulated by heat or injury. I hope this information is helpfu.

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Answer

Thank you for contacting us.

Yes, this product has been tested sucessfully in rabbit for Western Blotting. Our results show a clean band.

The antibody should specifically detect HSP70 and not HSP73.

Regarding local injury, I am not sure whether your levels will be elevated or not. Cells that are heat shocked do increase HSP70 so it seems reasonable that this would appear in your staining but this would have to be tested.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Vielen Dank für Ihre Antwort.

Die Frau Hoffmann ist außer Haus und ich hoffe es ist Ihnen Recht, dass ich mich um Ihren Fall kümmere.

Unser AbTrial ist nur ein Angebot, welches Sie nicht in Anspruch nehmen müssen. Natürlich können Sie jedes unserer Produkte bestellen dass Sie möchten ohne jegliche weitere Verpflichtung.

Ich hoffe dies hilft Ihnen weiter und ich wünsche Ihnen viel Glück mit Ihren Versuchen.

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Answer

Vielen Dank für Ihre Email.

Leider bieten wir generell keine Probegrößen zu Testzwecken an. Es ist logistisch bei über 95000 Produkten nicht möglich kleinere Größen anzubieten.

Unserem Wissen nach wurden ab47455 und ab79648 bisher noch nicht in Aspergillus getestet. Falls Sie dies selbst testen möchten, kann ich Ihnen zurzeit ein spezielles Angebot über einen 100%igen Abreview-Rabatt anbieten. Bei diesem Angebot bekommen Sie einen Rabatt für eine zukünftige Bestellung, wenn Sie uns ein Abreview mit dem Testresultat zusenden. Der Rabatt würde den ganzen Wert von ab47455 und ab79648 abdecken, und gegen eine erneute Bestellung weitere Produkte von uns verrechnet werden.

Um von diesem Angebot profitieren zu können, folgen Sie bitte diesen Schritten:

1.) Bestätigen Sie mir bitte, dass Sie ab47455 und ab79648 in Aspergillus testen möchten.

2.) Bitte bestellen Sie erst nach Erhalt der Rabattcodes.

3.) Bestellen Sie den Antikörper wie üblich per Telefon, Email oder Fax

4.) Testen Sie den Antikörper in Aspergillus.

5.) Teilen Sie uns das Ergebnis Ihres Tests durch ein Abreview mit und notieren Sie den Discount Code in dem Feld "additional notes"
Unter der folgenden URL können Sie mehr über unser Abreview System erfahren:
https://www.abcam.com/abreviews

6.) Die Rabattcodes sind nach dem Abschicken des Abreviews aktiv, und Sie können weitere Produkte zu dem gleichen Preis wie ab47455 und ab79648 bei uns bestellen (halten Sie bei der Bestellung bitte den Rabattcode). Bitte beachten Sie, dass der Rabattcode innerhalb von 4 Monaten nach Ausstellung eingelöst werden muss.

Der Rabattcode wird gültig unabhängig davon, ob Ihr Ergebnis positiv oder negativ ist.

Die Bedingungen zu unserem 100% Abreview Rabatt können Sie unter dem folgenden Link nachlesen:
https://www.abcam.com/collaborationdiscount

Ich hoffe, diese Informationen helfen Ihnen weiter. Falls Sie einen Rabattcode erhalten möchten oder weitere Fragen haben, zögern Sie bitte nicht, sich wieder an mich zu wenden.

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Answer

Thank you very much for your call yesterday and for your patience while I've been in touch with the lab about ab47455.

The antibody was used at a 1:100,000 dilution using a novel antibody amplification system, so these results are not typical and we recommend starting at a 1:1000 dilution.

I am sorry for the confusion, but I hope that this information is useful. Please let me know if you have any questions or if there is anything else that we can do for you.

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Answer

Thank you for getting back to me.

I agree that it does seem more sensible to try a different antibody as the vial you initially received was showing activity with the human protein. The two alternatives you have suggested, ab2787 or ab5439, have unfortunately not been tested in ELISA by us as yet and I would therefore not be certain of their performance using thisapplication.

Unfortunately, due to the size of our catalogue and the logistics involved, weare unable to provide sample sizes of antibodies. However, if you would like to try one of theseantibodies initially, I would be able to send you out a full vial to try, free of charge as a replacement forab47455 not working as expected. I would suggest that it may be beneficial to try performing a Western blot with your human and drosophila samples, just to check that the antibody is functioning as expected, prior to performing the ELISA with either of these antibodies.

Let me know if you would like me to send out one of these antibodies and which one you would like to try.Could you also confirmthat the addressI have for you is correct:



I look forward to receiving your reply.

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Answer

Thank you for contacting us.



I have asked our accounting department to issue a credit note for you, which can be redeemed against the invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question
Answer

Thank you for your reply.

Your credit note IDsare *******.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

The credit note ID is for your reference only and does not automatically guarantee the credit.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Answer

I am very pleased to hear you would like to accept our offer and test ab47544 in Bluegill tissues. This code will give you: 1 free PRIMARY ANTIBODY OR VALUE OFF OF ORDER before the expiration date. To redeem this offer, please submit an Abreview for Bluegill/Fish and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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1-10 of 17 Abreviews or Q&A

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