Overview

  • Product name

  • Description

    Rabbit polyclonal to Fbx32
  • Host species

    Rabbit
  • Specificity

    This antibody is only batch tested in WB and therefore we can only guarantee it for this application. Customers have successfully used this antibody in IHC and ICC but we have not tested it's performance in these applications. If you would like an antibody that is tested in ICC, we would recommend our recombinant antibody ab168372.
  • Tested applications

    Suitable for: WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human, Pig
    Predicted to work with: Chicken, Cow, Zebrafish
  • Immunogen

    Synthetic peptide within Human Fbx32 aa 300 to the C-terminus (internal sequence) conjugated to keyhole limpet haemocyanin. The exact sequence is proprietary.
    (Peptide available as ab74022)

  • Positive control

    • WB: Human, mouse and rat skeletal muscle tissue lysate. IHC-P: Human heart tissue. IHC-Fr: Rat muscle tissue. ICC/IF: Rat astrocyte cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab74023 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
WB Use a concentration of 2 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).

Target

Images

  • All lanes : Anti-Fbx32 antibody (ab74023) at 2 µg/ml

    Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 2 : Mouse skeletal muscle tissue lysate
    Lane 3 : Rat skeletal muscle tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa
    Additional bands at: 90 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 15 minutes
  • Paraffin-embedded human heart tissue stained for Fbx32 with ab74023 (1/2,000 dilution) in immunohistochemical analysis.

    Tissue underwent fixation in formaldehyde, peroxidase blocking, protein blocking and heat mediated antigen retrieval.  The secondary antibody was goat anti-rabbit conjugated to HRP.  For further experimental details please refer to abreview.

     

    See Abreview

  • Frozen sectioned rat muscle tissue stained for Fbx32 with ab74023 (1/200 dilution) (green) in immunohistochemical analysis.

    Tissue was fixed with 4% PFA overnight. Samples were then blocked with 10% donkey serum for 1 hour at 24°C followed by incubation with ab74023 for 24 hours at 4°C. A pig anti-rabbit Alexa Fluor488® conjugate was used as secondary antibody at a 1/1,000 dilution. Nuclei were stained using the standard Hoesch method and are depicted in blue.

     

     

    See Abreview

  • ab74023 staining Fbx32 in cultured rat astrocytes by ICC/IF at a 1/500 dilution (green).

    The cultured astrocytes were fixed with paraformaldehyde and blocked with 10% donkey serum for 30 minutes at 24°C. Cells were then stained with ab74023 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4.5 hours at 24°C. An Alexa Fluor-488® donkey anti-rabbit polyclonal antibody at 1/1,000 was used as the secondary antibody. Nuclei were stained with 1.43µM Hoechst and can be observed in blue.  Fbx32 expressed in the newly born astrocytes but does not co-localise with mature astrocytes.

    See Abreview

References

This product has been referenced in:

  • Ahmad N  et al. MicroRNA-672-5p Identified during Weaning Reverses Osteopenia and Sarcopenia in Ovariectomized Mice. Mol Ther Nucleic Acids 14:536-549 (2019). Read more (PubMed: 30769134) »
  • Cella PS  et al. Creatine supplementation in Walker-256 tumor-bearing rats prevents skeletal muscle atrophy by attenuating systemic inflammation and protein degradation signaling. Eur J Nutr N/A:N/A (2019). Read more (PubMed: 30806774) »
See all 11 Publications for this product

Customer reviews and Q&As

1-10 of 10 Abreviews or Q&A

Application
Western blot
Sample
Rat Cell lysate - whole cell (HEART)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
HEART
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 13 2019

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Non-reduced Denaturing (8% gel)
Sample
Pig Cell lysate - whole cell (smooth muscle cell)
Specification
smooth muscle cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Sep 29 2014

Application
Western blot
Sample
Rat Tissue lysate - whole (SD rat skeleta muscle tissue)
Loading amount
30 µg
Specification
SD rat skeleta muscle tissue
Treatment
30mg/Kg STZ IP injection
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 16 2013

Answer

Thank you for your reply.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab32845.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Answer

Thank you for contacting us.
I am sorry to hear the antibody is not performing as expected.

Please let me know if ab92281 would be a suitable replacement antibody, and I will arrange the shipment. Alternatively, a credit or a refund can be arranged.

I look forward to hear back from you, and resolve this case for you.

Read More

Answer

Thank you for your enquiry.

I would like to reassure you that our antibodies have all been tested successfully and would be covered by our guarantee in the applications listed on the datasheets. Therefore, any Fbx32 antibody tested in the species and application you are using should be suitable for your applications.

Also, although we produce many in-house antibodies, we obtain other antibodies from a wide range of sources. Therefore, comparison experiments will not often have been done.

If you wish to be more certain, I can suggest choosing the antibody that has been best characterized. By this I mean the antibody that has been tested in the most applications and has the most data available (on the datasheet). It is also helpful when running WB to know that the blocking peptide is available, as a blocking control may be invaluable in interpreting your data.

Ab74023 has been tested in more applications and does have the associated peptide available.


I hope you have found this information helpful. Should you have any further questions please do not hesitate to ask.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Astrocytes culture)
Specification
Astrocytes culture
Fixative
Paraformaldehyde
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Feb 10 2012

Answer

Thank you for contacting us. The antibody ab74023 will be useful for IHC, but we do not know if it has equal affinity for free Fbx32 and the same protein bound to substrate or ubiquitin. The antibody is raised against a peptide from the C-terminus of Fbx32. https://www.abcam.com/Fbx32-antibody-ab74023.html If you are in Hong Kong, then I recommend sending an e-mail to hk.orders@abcam.com. That office will be happy to assist you with ordering. If you hospital is somewhere else, for instance Guangdong, I recommend contacting our distributor, Guangzhou Genewindows. Their URL is http://www.genewindows.com . Other distributors can be found by going to the datasheet for ab74023 (or any other antibody) and selecting your region from the distributor details menu.

Read More
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Muscle)
Specification
Muscle
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% TritonX in 0.1% PBS
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Jun 02 2011

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Heart)
Specification
Heart
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako Target Retrieval solution
Permeabilization
No
Blocking step
5 minutes of peroxidase block then 10 minutes of protein block. These are ready-to-use reagents purchased from Dako as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 20°C

Mr. Antibody Solutions

Verified customer

Submitted May 05 2010

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