Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

Reviews (6)Q&A (31)References (306)

Overview

  • Product name

    Rabbit Anti-Mouse IgG H&L (HRP)
    See all IgG secondary antibodies

  • Description

    Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP)

  • Host species

    Rabbit

  • Target species

    Mouse

  • Tested applications

    Suitable for: Dot blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, ICC/IF, WBmore details

  • Immunogen

    Mouse IgG whole molecule

  • Conjugation

    HRP

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Gentamicin sulphate
    Constituents: 0.42% Potassium phosphate, 0.87% Sodium chloride, 1% BSA
  • Concentration information loading...

  • Purity

    Affinity purified

  • Purification notes

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Mouse IgG coupled to agarose beads.

  • Conjugation notes

    Horseradish Peroxidase (HRP)

  • Clonality

    Polyclonal

  • Isotype

    IgG

  • General notes

    Many of our customers have reported seeing brown precipitates in the vials. The brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. Our customer’s feedback says the antibody worked great. If in case the antibody fails to give results then please contact our Scientific Support team for assistance.

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab6728 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
Dot blot Use at an assay dependent dilution.
ELISA 1/1000.
IHC-P Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution.
Immunomicroscopy Use at an assay dependent dilution.
ICC/IF 1/1000 - 1/5000.
WB 1/2000 - 1/10000. PubMed: 17200442

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)Gao et al PLoS One. 2016 Apr 28;11(4):e0154419. doi: 10.1371/journal.pone.0154419. eCollection 2016. Fig2. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Distribution, expression and correlation of IL-22 cells with liver fibrosis in the liver.

    Immunohistochemistry double staining identities IL-22 cells were produced by CD4 cells in liver tissue (Panel B).

    Black arrows indicate double-positive cells.

    Paraffin-embedded, formalin-fixed human liver tissues were incubated with anti-IL-22 (ab18499) or α-SMA antibody (ab7817) overnight at 4°C followed by heat mediated antigen retrieval with sodium citrate buffer (pH 6.0, 0.1 mol/L) for 20mins and blocking endogenous peroxidase activity with 0.3% H2O2. Revelation of primary antibody was carried out using horseradish peroxidase (HRP)-conjugated rabbit polyclonal anti-mouse (ab6728), secondary antibody followed by diamino-benzidine (DAB) and haematoxylin coloration. Double staining was performed with 3-amino-9-ethyl-carbazole (red color) for IL-22 and BCIP/ NBT (indigo color) for CD4.

    Positively stained cells were counted at high-power field (hpf, ×400) according to described protocols.

  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
    Immunohistochemistry (PFA perfusion fixed frozen sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

    ab6728 was used at dilution 1/500 with the primary antibody ab3089 in IHC-Fr (PFA perfusion). See the review on ab3089.

  • Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
    Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

    ab6728 was used at dilution 1/2000 with the primary antibody ab72009 in WB. See the review on ab72009.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)This image is courtesy of an anonymous Abreview.

    ab81785 staining LC3A/B in Mouse kidney tissue sections by Immunohistochemistry (IHC-P - formaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 5% protein block for 5 minutes at room temperature; antigen retrieval was by heat mediation. Samples were incubated with primary antibody (1/50) for 1 hour. Anti-mouse HRP (ab6728) (1/200) was used as the secondary antibody.

  • ELISA - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
    ELISA - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

    ab6728 was used at dilution 1/1000 with the primary antibody ab11038 in ELISA. See the review on ab11038.

References

This product has been referenced in:

  • Jiang L  et al. Inhibition of microRNA-103 attenuates inflammation and endoplasmic reticulum stress in atherosclerosis through disrupting the PTEN-mediated MAPK signaling. J Cell Physiol 235:380-393 (2020). Read more (PubMed: 31232476) »
  • Tian BY  et al. Specific knockdown of WNT8b expression protects against phosphate-induced calcification in vascular smooth muscle cells by inhibiting the Wnt-ß-catenin signaling pathway. J Cell Physiol 234:3469-3477 (2019). Read more (PubMed: 30461014) »
See all 306 Publications for this product

Publishing research using ab6728? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-10 of 37 Abreviews or Q&A

Very sensitive and specific secondary antibody

 Excellent
Abreviews
abreview image
Application
Western blot
The dilution used for this secondary antibody was 1/20000 and we obtained intesnse specific bands even at very short incubation times. It works very well with phosphorylated proteins which sometimes are harder to detect (in these cases we use 1/10000 dilution or 1/5000).

Figure legend
Western blot image of actin bands. HCT116 cells transfected with non-targeting siRNA (lane #1) or CDK4/6 siRNA (lane #2).

Abcam user community

Verified customer

Submitted Oct 02 2018

Immunoblot detection of B-3-Tubulin

 Excellent
Abreviews
abreview image
Application
Western blot
20 μg of micro tubular protein from SH-SY5y cells were used to detect B-3-Tubulin (Primary antibody - 1:500 dilution overnight, Secondary antibody (ab6728 - 1: 2000 dilution for 1h).

Abcam user community

Verified customer

Submitted Feb 28 2018

Excellent 2nd antibody for WB

 Excellent
Abreviews
abreview image
Application
Western blot
A excellent 2nd Ab for WB (1:2,000 dilution) used with ab9484 (1:10000)

Mr. Baptiste Jude

Verified customer

Submitted Oct 10 2016

This is a good 2nd antibody

 Excellent
Abreviews
abreview image
Application
Western blot
I used this antibody, when I perform WB.
This antibody is very sensitive.
Dilution concentration that I used is 1/2000 in TBST(5% BSA) for 1hour.

Abcam user community

Verified customer

Submitted May 30 2014

The product is good

 Excellent
Abreviews
Application
Western blot
When I perform WB, that is very sensitive. making band is clearly

Abcam user community

Verified customer

Submitted Jan 23 2014

good for mouse primary antibody

 Excellent
Abreviews
abreview image
Application
Western blot
strong detection

Abcam user community

Verified customer

Submitted Jul 11 2013

Question

Our customer purchased ab6728 when we had promotion last December and opened and used recently. It showed high background so the customer tried several times but failed to optimize the condition.
So, the customer tried other brand’s(GenDEPOT) secondary antibody and finally, he got good results. I have attached file and you can see the data.
I explained about abpromise but the customer would like to ask your opinion and recommendation about his data.
Please check attached file.

Best regards

Read More
Answer

Thank you for your email. I am sorry to hear that customer have been experiencing problems with this antibody.

I have read the details you have kindly provided and have following further suggestions for troubleshooting;

- Please try 5% BSA for blocking with 2 hours incubation
- Heat the samples for 10 minutes 100C
- Try no primary control

It is true that the antibody is out of guarantee period so I am sorry we cannot provide replacement products; we can only provde the troubleshooting suggestions. If however this customer is your loyal customer then you can certainly offer them incentives for keeping them on board.

I hope this information will be helpful. SHould you have any question please do not hesitate to contact me.

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Question

ecd a problem with our product, which including mouse anti His Tag (4D11) cat no. AB5000 (lot GR44936-1) and AB6728 (Lot GR15565-6) and wants to claim this item under the abpromise guarantee. Her detailed information about her condition using to detect the protein is in the attached file.

However, I’m not sure if it’s possible to help her to optimize the reaction and get a better result. Because her comment is that primary antibody cannot bind to her target protein at all.

It would be my appreciate if you are able to help me to solve her problem?

Read More
Answer

Thank you for contacting us. I am sorry to hear these antibodies are not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. 1) About the secondary antibody ab6728: as indicated on the datasheet, many of our customers have reported seeing brown precipitates in the vials. The brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. However, I would be pleased to replace, if you wish, this antibody with an alternative catalog reference : ab97046, www.abcam.com/ab97046. 2) About the anti-His tag ab5000: to our knowledge this antibody has never been tested for one hour incubation only but overnight at 4ºC. Some antibodies bind better to their target at 4ºC, some other at room temperature. I think it would be worth trying an overnight incubation at 4ºC for this antibody. The blocking solution should not be problem, this antibody works fine in milk as well as in BSA. Please let me know about the results after an overnight incubation at 4ºC. If the antibody is still not working I will be pleased to offer a replacement or a credit note. Thank you for your cooperation. I look forward to hearing from you with your results using ab5000 and with details of how you would like to proceed with ab6728.

Read More

Question

Dear Technical Team,

Hope this e-mail finds you well!

Kindly suggest the exact concentration of Rabbit polyclonal Secondary Antibody to Mouse IgG - H+L (HRP) (ab6728) since on it's datasheet the concentration is mentioned as 1 mg at 2mg/ml .

Looking forward towards your valued response on it.

With Best Regards,

Read More
Answer

Thank you for your message.

I am pleased to help answer your question.

I can confirm that '1 mg at 2mg/ml' means that the vial contains 1 mg of antibody at a concentration of 2 mg.ml. The concentration is 2 mg/ml. (Therefore the vial contains 500 ul).

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Question

I'm confused because when I had inquiry about the precipitation before, I received emailusing them with gentle vortex but,
this time, you answered spin down it and use only supernatant.
Let me know which method is right.


Best regards,

Read More
Answer

Thank you for your reply.

I can confirm that for the HRP conjugated antibodies the approach that Irecommended is the correct one. Spining the vial down will prevent free HRP to bind to un-specifically to samples and therefore avoid increased background.

If the antibody is not purified and therefore also not conjugated then we would recommend gentle vortexing. This could be whole antiserum for example.
In this casethe antibody does contain proteins and other solutes it would not be abnormal to see a little cloudiness or precipitation, especially after being stored at a low temperature and/or for a long period of time. Most of the time bringing the antibody up to room temperature, mixing it well and then diluting it to a working concentration will help incorporate any protein aggregates and precipitation.

I hope this information will explain the different suggestions we gave and I apologise for the confusion.

Read More

1-10 of 37 Abreviews or Q&A

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