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:
  • Zhu X  et al. The role of calpains in ventilator-induced diaphragm atrophy. Intensive Care Med Exp 5:14 (2017). WB ; Rat . Read more (PubMed: 28290154) »
  • Li F  et al. Ginsenoside Rg1 prevents starvation-induced muscle protein degradation via regulation of AKT/mTOR/FoxO signaling in C2C12 myotubes. Exp Ther Med 14:1241-1247 (2017). WB ; Mouse . Read more (PubMed: 28781621) »

See all 7 Publications for this product

Customer reviews and Q&As

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
Username

Abcam user community

Verified customer

Submitted Sep 29 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

Submitted Apr 16 2013

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.
Read More

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 ...

Read More

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 ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Feb 10 2012

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 o...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Jun 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Mr. Antibody Solutions

Verified customer

Submitted May 05 2010

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

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