Overview

  • Product name
    Anti-4 Hydroxynonenal antibody [HNEJ-2]
    See all 4 Hydroxynonenal primary antibodies
  • Description
    Mouse monoclonal [HNEJ-2] to 4 Hydroxynonenal
  • Host species
    Mouse
  • Tested applications
    Suitable for: WB, IHC-P, Flow Cyt, ELISA, IHC-FoFrmore details
  • Immunogen

    Chemical/Small Molecule: 4-Hydroxy-2-Nonenal modified KLH.

  • Positive control
    • Colorectal carcinoma cells.
  • General notes

    This product was changed from ascites to tissue culture supernatant on 28th February 2018. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab48506 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 - 15 µg/ml.
IHC-P Use a concentration of 10 - 20 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol. But NOTE: Fixation using Bouin's solution is recommended. In this case, antigen retrieval process is not required.
Flow Cyt Use at an assay dependent concentration.

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

ELISA Use at an assay dependent concentration. PubMed: 22881716
IHC-FoFr Use at an assay dependent concentration.

Target

  • Relevance
    Aldehydic products of lipid peroxidation, such as 4 hydroxynonenal (4 HNE), have been implicated in the etiology of pathological changes under oxidative stress as a key mediator of oxidative stress induced cell death. It is a stable product of lipid peroxidation, is proarrhythmic and may contribute to the cytotoxic effects of oxidative stress.
  • Cellular localization
    Cytoplasmic
  • Alternative names
    • 4 HNE antibody
    • 4-Hydroxy-2-Nonenal antibody

Images

  • ab48506 at a 1/25 dilution staining 4-Hydroxy-2-Nonenal in mouse heart tissue sections by Immunohistochemistry (paraffin embedded) incubated for 15 hours at +4°C. Fixed with formaldehyde, heat mediated antigen retrieval step performed using citrate buffer. Blocked using 5% serum for 20 minutes at 20°C. Secondary used undiluted polyclonal Goat anti-mouse IgG conjugated to Alexa Fluor 594.

    See Abreview

  • ab15463 staining 4-Hydroxynonenal in the Mouse melanoma cancer tissue sections by IHC-FoFr (PFA perfusion fixed frozen Sections). Tissue samples were fixed with 4% Formalin, Antigen retrieval carried out heat mediation using anautoclave for 10 minutes, permeabilized using 0.1% SDS and blocked with 1% BSAfor 15 minutes at 25°C. The sample was incubated with primary antibody (1/25 in PBS + 1% BSA) at 4°C for 12 hours.An Alexa Fluor®488-conjugated Goat anti-mouse IgG(H+L) monoclonal(1/100) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Lee HL  et al. Yuk-Mi-Jihwang-Tang , a Traditional Korean Multiple Herbal Formulae, Improves Hippocampal Memory on Scopolamine Injection-Induced Amnesia Model of C57BL/6 Mice. Evid Based Complement Alternat Med 2018:2821040 (2018). Read more (PubMed: 29507588) »
  • Yan Z  et al. Curcumin exerts a protective effect against premature ovarian failure in mice. J Mol Endocrinol 60:261-271 (2018). Read more (PubMed: 29437881) »
See all 64 Publications for this product

Customer reviews and Q&As

1-10 of 27 Abreviews or Q&A

Application
Western blot
Sample
Mouse Tissue lysate - whole (Liver)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (8%)
Loading amount
35 µg
Treatment
5% ethanol for 10 days
Specification
Liver
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Ms. Ji Hye Han

Verified customer

Submitted Oct 23 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Liver)
Antigen retrieval step
None
Permeabilization
No
Specification
Liver
Blocking step
Vector Lab M.O.M. blocking agent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.8% · Temperature: 25°C
Fixative
Bouin's fixative

Abcam user community

Verified customer

Submitted Aug 19 2016

Question
Answer

ab48506 antibody will detect only 4-HNE modified proteins.

This antibody is specific for 4-HNE-lysine, 4-HNE-histidine and
4-HNE-cysteine adduct.
By inhibition test, this antibody has a much higher affinity for the
4-HNE-histidine adduct than 4-HNE-lysine or 4-HNE cysteine adduct.

ab48506 shows almost negligible reactivity with proteins that were
treated with other aldehydes, such as 2-nonenal, 2-hexenal, 1-hexanal,
4-hydroxy-2-hexenal, formaldehyde, or glutaraldehyde.

Read More

Answer

Gracias por contactarnos.


No especificamos las especies para las que este anticuerpo está recomendado pues el antígeno corresponde a una estructura química, una molécula relativamente pequeña que es constante en todas las especies.

Siempre y cuando se exprese en el tejido que el cliente quiere usar, no habría problema en probarlo con secciones de ratón.

Tened en cuenta que en nuestro laboratorio se ha validado en secciones embebidas en parafina y IHC-FoFr.

Si tenéis cualquier otra consulta, no dudes en contactarnos de nuevo.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Melanoma cancer)
Specification
Melanoma cancer
Fixative
4% Formalin
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: autoclaved for 10min
Permeabilization
Yes - 0.1% SDS
Blocking step
BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 25°C

Ms. Seontae Kim

Verified customer

Submitted Dec 19 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (B16F10 melanoma)
Loading amount
30 µg
Specification
B16F10 melanoma
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Ms. Seontae Kim

Verified customer

Submitted Dec 18 2012

Answer

Thank you for contacting us.

For IHC-P we recommend to dilute the anti-4 Hydroxynonenal antibody [HNEJ-2] ab48506 in TBS with 1% BSA at a concentration of 10 - 20 µg/ml.
As indicated on the product datasheet, a heat mediated antigen retrieval via the microwave method can be performed before commencing with IHC staining protocol. But a fixation of the samples using Bouin's solution is recommended. In this case, antigen retrieval process is not required.

The use of Dako products should be fine, however we have no such evidence for this particular antibody.

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

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Answer

Sorry, I neglectedadd that the mouse monoclonal, ab48506,has also been used in IHC in the following publications:

Li J et al. Up-regulation of p27kip1 contributes to Nrf2-mediated protection against angiotensin II-induced cardiac hypertrophy. Cardiovasc Res 90:315-24 (2011). IHC-P. PubMed: 21245055

Kumar A et al. Resveratrol restores nrf2 level and prevents ethanol-induced toxic effects in the cerebellum of a rodent model of fetal alcohol spectrum disorders. Mol Pharmacol 80:446-57 (2011). IHC. PubMed: 21697273

Li J et al. Nrf2 protects against maladaptive cardiac responses to hemodynamic stress. Arterioscler Thromb Vasc Biol 29:1843-50 (2009). IHC-P. PubMed: 19592468

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

Read More

Answer

Thank you for contacting us and your interest in our products.

The Anti-4 Hydroxynonenal antibody (ab46545) has been reported to work well for one of our customers in using the antibody in rat tissue sections. They found that using this antibody showed that the perfusion of H2O2 increased the level of 4-HNE from the baseline condition. High-resolution, confocal imaging showed the increase of 4-HNE labeling at the endothelial cell surface with H2O2 exposure.

This was performed using the following protocol:

https://www.abcam.com/index.html?datasheet=46545&tab=abreviews&intabreviewid=26369

We also have an alternative antibody (mouse monoclonal https://www.abcam.com/4-Hydroxynonenal-antibody-HNEJ-2-ab48506.html) to the same target which has also shown good staining of mouse heart tissuewhen used in IHC:

https://www.abcam.com/index.html?datasheet=48506&tab=abreviews&intabreviewid=16917

Both of these antibodies would be covered by the Abrpomise guarantee if used in IHC of paraffin embedded sections. More information on the Abpromise guarantee can be found from the following link:

https://www.abcam.com/abpromise

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

Read More

Answer

Thanks for sending along the document. I have forwarded it to our Accounts department. I will let you know what they say as soon as possible.

Please feel free tocontact me in the meantime if you have any other questions.

Read More

1-10 of 27 Abreviews or Q&A

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

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