Overview

  • Product name
  • Description
    Rabbit polyclonal to Hsp90
  • Host species
    Rabbit
  • Specificity
    Detects 90kD proteins corresponding to the molecular mass of hsp90aß.
  • Tested applications
    Suitable for: ELISA, IP, IHC-P, IHC-Fr, ICC/IF, ICC, Flow Cyt, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Xenopus laevis
  • Immunogen

    Human Hsp90.

  • Positive control
    • RAW 264.7 whole cell lysate (ab7187) can be used as a positive control in WB. HeLa Cell Lysate (Heat Shocked). Hsp90 Protein.

Properties

Applications

Our Abpromise guarantee covers the use of ab13495 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
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.
Flow Cyt 1/250.

(see Abreview).

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/20000 - 1/40000. Detects a band of approximately 90 kDa (predicted molecular weight: 83.2 (beta) , 84.5 (alpha) kDa).

Target

  • Function
    Molecular chaperone that promotes the maturation, structural maintenance and proper regulation of specific target proteins involved for instance in cell cycle control and signal transduction. Undergoes a functional cycle that is linked to its ATPase activity. This cycle probably induces conformational changes in the client proteins, thereby causing their activation. Interacts dynamically with various co-chaperones that modulate its substrate recognition, ATPase cycle and chaperone function.
  • Sequence similarities
    Belongs to the heat shock protein 90 family.
  • Domain
    The TPR repeat-binding motif mediates interaction with TPR repeat-containing proteins like the co-chaperone STUB1.
  • Post-translational
    modifications
    ISGylated.
    S-nitrosylated; negatively regulates the ATPase activity and the activation of eNOS by HSP90AA1.
  • Cellular localization
    Cytoplasm. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • Heat shock 86 kDa antibody
    • Heat shock protein 90kDa alpha cytosolic class A member 1 antibody
    • Heat shock protein 90kDa alpha cytosolic class B member 1 antibody
    • Heat shock protein HSP 90 alpha antibody
    • Heat shock protein HSP 90 beta antibody
    • Heat shock protein HSP 90-alpha antibody
    • HS90A_HUMAN antibody
    • HSP 84 antibody
    • HSP 86 antibody
    • Hsp 90 antibody
    • HSP86 antibody
    • HSP90A antibody
    • HSP90AA1 antibody
    • HSP90AB1 antibody
    • HSP90B antibody
    • HSPC1 antibody
    • HSPC2 antibody
    • HSPCAL1 antibody
    • HSPCAL4 antibody
    • Renal carcinoma antigen NY-REN-38 antibody
    see all

Images

  • All lanes : Anti-Hsp90 antibody (ab13495) at 1/10000 dilution (2 hours at room temperature)

    Lane 1 : A431 with BSA for 30 minutes at room temperature
    Lane 2 : A549 with BSA for 30 minutes at room temperature
    Lane 3 : HCT116 with BSA for 30 minutes at room temperature
    Lane 4 : Hela with BSA for 30 minutes at room temperature
    Lane 5 : HEK293 with BSA for 30 minutes at room temperature
    Lane 6 : HepG2 with BSA for 30 minutes at room temperature
    Lane 7 : HL-60 with BSA for 30 minutes at room temperature
    Lane 8 : HUVEC with BSA for 30 minutes at room temperature
    Lane 9 : Jurkat with BSA for 30 minutes at room temperature
    Lane 10 : MCF7 with BSA for 30 minutes at room temperature
    Lane 11 : PC3 with BSA for 30 minutes at room temperature
    Lane 12 : T98G with BSA for 30 minutes at room temperature

    Lysates/proteins at 2 µg per lane.

    Blocking peptides at 1.5 % per lane.

    Secondary
    All lanes : HRP Donkey anti-rabbit IgG, 1 hour at room temperature

    Predicted band size: 83.2 (beta) , 84.5 (alpha) kDa

  • ab13495 staining Hsp90 in murine RAW 264 (ab7187).7 cells by Immunocytochemistry/ Immunofluorescence.Cells were fixed in paraformaldehyde, permeabilized using 0.1% Triton-X100 in 2% BSA for 15 minutes, blocked with 2% BSA for 1 hour at 4°C and then incubated with ab13495 at a 1/150 dilution. The secondary used was an Alexa-Fluor 488 conjugated chicken anti-rabbit IgG (H+L) used at a 1/500 dilution.

    See Abreview

  • ab13495 staining Hsp90 in Mouse backskin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with Bouin's fixative solution. Samples were incubated with primary antibody (1/100) for 1 hour at room temperature. A FITC-conjugated Goat anti-rabbit IgG polyclonal (1/50) was used as the secondary antibody.

  • ab13495 at a 1/100 dilution staining Hsp90 in human PMN cells by Immunocytochemistry/ Immunofluorescence incubated for 4 hours at 37°C. PFA fixed. Blocked using 2% BSA for 1 hour at 22°C. Secondary used at 1/250 polyclonal Goat anti-rabbit IgG (H+L) conjugated to Alexa Fluor 568 (ab175471).Left image: DAPI staining nuclei (blue)Middle image: Hsp90 (red)Right image: Overlay

    See Abreview

  • ab13495 staining Hsp90 in Human platelet cells by Flow cytometry.
    Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/250 dilution and incubated for 18 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.

    P : Permeabilized; US : Unstained, Red Peak; IgG RB : IgG Rabbit (Isotype Control) ab171870), Blue Peak; HSP90, Green peak.

    See Abreview

  • Anti-Hsp90 antibody (ab13495) at 1/1000 dilution + whole cell lysate prepared from human platelets treated with A23187 for 1 hour at 20 µg

    Secondary
    HRP conjugated goat anti-rabbit polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Predicted band size: 83.2 (beta) , 84.5 (alpha) kDa
    Observed band size: 90 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 36 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 5 minutes

    See Abreview

References

This product has been referenced in:
  • Cho S  et al. Progerin phosphorylation in interphase is lower and less mechanosensitive than lamin-A,C in iPS-derived mesenchymal stem cells. Nucleus 9:230-245 (2018). Read more (PubMed: 29619860) »
  • An T  et al. Comparative analysis of proteomes between diabetic and normal human sperm: Insights into the effects of diabetes on male reproduction based on the regulation of mitochondria-related proteins. Mol Reprod Dev 85:7-16 (2018). Read more (PubMed: 29149484) »
See all 23 Publications for this product

Customer reviews and Q&As

1-7 of 7 Q&A

Answer

Thank you for providing those details.

The sequences provided are only truncations of the proteins and I have therefore not been able to compare them to the immunogens used to raise the antibodies ab2787 and ab13495. I cannot therefore ascertain how likely it is that the antibodies will cross react with your customers bufo bankorensis samples.

I have however performed a blast of the two immunogens used and they each resulted with a few sequences of toad derived proteins matching with higher than 85% homology. It is therefore possible that the antibodies will cross react. I am therefore willing to provide the testing discount codes as a gesture of goodwill. The codes are as follows:

ab2781, Discount code: xxxxx expiration date 26th December 2012
ab13495, Discount code: xxxxx expiration date 26th December 2012

I hope this has been of help. If you have any further questions, please do not hesitate to contact us again.

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Answer

Thank you very much for providing those sequences.

In order to help the customer, could you please provide me the sequence in text format (i.e. in the body of the email, or as a word document). This will allow me to copy and paste the sequence to be able to find the protein sequence and align this to the immunogens used to raise the Hsp90 and Hsp70. This will allow me to advise the customer as to whether the antibodies are likely to be able tocross-react withthe proteins of interest or not.

I look forward to receiving your reply.

Read More

Answer

Thank you for contacting us.

I have been unable to find a sequence for Hsp90 and Hsp70 in bufo bankorensis, I am therefore not able to say if the antibodies are likely to be able to detect the proteins from these samples. Would you be able to ask the customer is they have the protein sequences for Hsp90 and Hsp70from bufo bankorensis and I will check this for them. I can then let you know if the customer would be eligiblefor the testing discount scheme or not.

Many thanks in advance for your assistance in this matter.

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Answer

Thank you for submitting your Abreview and for your honest feedback.
As you had rated your experience in the review as 'Below average', I just wanted to follow up with you to get some more details as to what you were unhappy about regarding use of ab62817 in your ICC experiiment.
Thanks in advance for the additional information.

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Answer

Thank you for contacting us with your question about ab13495. As this Hsp90 antibody has not been purified and is provided as whole antiserum, we do not know the specific IgG concentration. We estimate that the specific antibody concentration in whole antiserum is less than 0.5 mg/mL. For IP, we recommend using whole antiserum antibodies at a dilution between 1:50 and 1:100 in IP, so you may want to try a couple dilutions within this range in order to optimize the dilution to your samples. I hope this information will be useful, but please let me know if you have any further questions and I'll be happy to help.

Read More

Answer

Thank you for sending details regarding your protocol. At this point i would like to make the following suggestions. Try incubating with the primary antibody for overnight at 4C. Ensure that the protein transfered properly to the membrane (you can check this with Ponceaue S). Also, you mentioned that your secondary antibody was "goat antibody raised in rabbit IgG." As this primary antibody was raised in a rabbit, the secondary antibody must be anti-rabbit IgG. For example, goat polyclonal to rabbit IgG (raised in a goat). Please check to make sure that your secondary antibody is compatible for use with ab13495. I hope this helps. Please contact us again if you need additional assistance with this antibody.

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Question
Answer

Thank you for your email. Yes, it is recommended to use ab13495 at a dilution of 1:35,000 in Western blotting. This dilution was found to be sufficient to detect Hsp90 in HeLa cell lysate. If you have any further questions, please let us know.

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