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

Reviews (8)Q&A (31)References (654)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
  • Immunohistochemistry (PFA perfusion fixed frozen sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
  • Western blot - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)
  • ELISA - Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

Key features and details

  • Rabbit Anti-Mouse IgG H&L (HRP)
  • Conjugation: HRP
  • Host species: Rabbit
  • Isotype: IgG
  • Suitable for: Dot blot, ELISA, IHC-P, IHC-Fr, Immunomicroscopy, ICC/IF, WB

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase Biotin FITC Texas Red ® TRITC Unconjugated Unconjugated

Overview

  • Product name

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

  • 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

    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

The Abpromise guarantee

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 (8)
1/2000 - 1/10000. PubMed: 17200442
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.

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (654)

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

ab6728 has been referenced in 654 publications.

  • Zhou C  et al. Exosome-derived miR-142-5p remodels lymphatic vessels and induces IDO to promote immune privilege in the tumour microenvironment. Cell Death Differ 28:715-729 (2021). PubMed: 32929219
  • Song T  et al. Zearalenone exposure affects the Wnt/ß-catenin signaling pathway and related genes of porcine endometrial epithelial cells in vitro. Anim Biosci 34:993-1005 (2021). PubMed: 32898953
  • Qiu J  et al. Role of TGF-ß1-mediated epithelial-mesenchymal transition in the pathogenesis of tympanosclerosis. Exp Ther Med 21:6 (2021). PubMed: 33235615
  • Liu X  et al. The AMPK-mTOR axis requires increased MALAT1 expression for promoting granulosa cell proliferation in endometriosis. Exp Ther Med 21:21 (2021). PubMed: 33235630
  • Kashkoulinejad-Kouhi T  et al. Enhancement of cisplatin sensitivity in human breast cancer MCF-7 cell line through BiP and 14-3-3? co-knockdown. Oncol Rep 45:665-679 (2021). PubMed: 33416155
View all Publications for this product

Customer reviews and Q&As

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21-30 of 39 Abreviews or Q&A

Question

Dear Technical Please issue coc for ab8249 from lot GR41773-1 And ab6728 from lot GR15565-7 Please don't forget to mention expiry date Best regards  

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Answer

Thank you for your enquiry. Please find attached the CoCs you requested. We do not have set expiration dates for our products. Most antibodies are stable and can last for anywhere from a few months to several years if stored properly, so we strongly recommend that you follow the storage instructions on the datasheet for the antibody you purchased. These conditions will vary among our antibodies, therefore, it is important to verify the storage conditions for each of our products when you receive them. We guarantee all of our products to work for at least 6 months from the date of purchase when stored correctly. For more information on antibody storage and stability, please visit our Antibody Storage Guide on our Protocols and Troubleshooting Tips page (https://www.abcam.com/index.html?pageconfig=popular_protocols). I hope this information helps. Please do not hesitate to contact us if you need anything further.

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Question

      There is a quality issue for the antibody ab6728. Please check attachment.   We request replacement.    

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Answer

Thank you for contacting us and for letting us know about this precipitate. The antibody ab6728 does often contain a brown precipitate, and it should perform as expected. We encourage you to try this antibody and let us know if there are any problems with the results. However, if you still have concerns and do not wish to try this vial, I will be happy to send a replacement vial of ab6728 or a different anti-mouse secondary antibody. Please keep me updated of the results with this vial, or let me know if you would prefer the replacement. I apologize for any inconvenience this issue has caused.

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Question

Phone call regarding precipitate formation in Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) ab6728.

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Answer

I have looked into your report of precipitation of the secondary HRP antibody you have received, ab6728. I am pleased to inform you that this is normal with this antibody due to the HRP conjugation and should not affect the activity of the antibody. I would recommend gently vortexing and using as normal. However, if you find that the antibody does not perform as expected please let us know and we will be happy to offer a free of charge replacement or refund under our Abpromise guarantee. More information of this can be found here: https://www.abcam.com/index.html?pageconfig=resource&rid=10372 If you have any further questions please do not hesitate to contact us again.

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Question

Rabbit anti mouse IgG HRP ordered from Abcam company is having some clot in the vial and it is unused

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Answer

Thank you very much for informing us about this problem of the customer. Indeed, this HRP conjugated antibody is known for potentially having precipitates inside. This is due to the HRP conjugation and won't affect the performance of the antibody. We had recently followed up with customers who had this precipitate and tried the antibody. They told us that the antibody was working well. I do suggest therefore, if the clot the customer is seeing is brownish and looks like a precipitate, than the customer can try this antibody as usual. If there is nevertheless a problem with the results, please advise us and we will immediately replace the product. Thank you very much for your help. Please let me know if you have any questions.

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Question

We have concerns regarding the brown precipitate seen in the vial.

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Answer

Thank you for contacting us. Many of our customers have reported similar issues to us. We are completely aware of this problem. We have followed the complaints to many of our customers; their feedback says the antibody worked great, they found no problem at all. Please be advised that the brown precipitates are very common with HRP conjugated antibodies; we suggest vortexing the vial and using this antibody as normal. If in case the antibody fails we will be happy to offer a free of charge replacement under our Abpromise guarantee. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Question

BATCH NUMBER 212428 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM I purchased rabbit anti-mouse antibody(ab6728, secondary antibody). I tried to western using human plasma including negative control. I diluted 1/2000, 1/5000, and 1/30,000. However I always detected Ig heavy and light chain in negative blot. It seems like rabbit antibody(Ig) is cross-reacting with human IgG. SAMPLE Human Plasma PRIMARY ANTIBODY fibronectin (ab26245): 1/100, 1/500, 1/1000 Trx (ab1754):1/100, 1/150 Overnight Incubation at 4 degree or 1hr incubation in RT TBS-Tween 20(0.05%)was dilution buffer and washing buffer DETECTION METHOD Immobilon (Millipore), Femto (piearce) POSITIVE AND NEGATIVE CONTROLS USED negative control : was incubated with only 2nd antibody (ab6728) ANTIBODY STORAGE CONDITIONS aliquted and stored -20 degree. SAMPLE PREPARATION 5 ul of diluted plasma was added with sds-loading buffer and heated for 4 min. and then loaded into well. AMOUNT OF PROTEIN LOADED 20 ug of plasma ELECTROPHORESIS/GEL CONDITIONS 4~12%, 3~8%, 12% reducing sds-gel TRANSFER AND BLOCKING CONDITIONS 13 mM Tris, 81 mM Glycine, 10% Methol-> transfer buffer 2 hours 5% skin milk, overnight blocking at 4 degree SECONDARY ANTIBODY ab 6728 1 hour incubation in RT TBS-Tween 20(0.05%)was dilution buffer and washing buffer HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? >10 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

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Answer

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. I have read your technical questionaire and I have a few comments. The results that you have been obtaining are indeed concerning. You are detecting two bands most likely human heavy and light IgG. Please can you tell me whether these bands are of an appropriate size for this conclusion or whether the reactivity could be attributable to something else. I look forward to hearing from you.

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Question

I purchased the antibody-ab3364 and ab6728 from your company.and I have used it to do my experiment,but the result is not good. I have no reference.So I have three questions to consult you.and hope to get your answeras fast as possible.Thanks a lot.NO1.Sections should be pre-treated with an antigen unmasking solution.Could you tell me the details and what this antigen unmasking solution is and what to do step by step. NO2.Could you give me the result (you did )of P-glycoprotein IHC-P and WB using this antibody. so I can reference and contrast. I use rat brain to do my experiment so the result of this tissue is the best.NO3. if it existed repeated freeze/thaw cycles, how many times ab6728 will become invalid. thank you very much .I am looking forward to getting your answer as soon as possible.

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Answer

Thank you for your enquiry. The antigen retrieval method for this antibody is heat-induced epitope retrieval at a high pH (EDTA, pH 8-9). You may find a detailed protocol at IHCWorld.com: http://www.ihcworld.com/_protocols/epitope_retrieval/tris_edta.htm Unfortunately I do not have an image of this antibody used in Western blotting or IHC. I do not see that the antibody was tested in brain tissue, but human liver showed positive staining along luminal surfaces of bile canaliculi, and human colon showed positive staining on the luminal surface of secretory epithelium. In Western blot, the band size is predicted to be 141kDa, but may run slightly higher due to glycosylation. I do not know how many freeze/thaw cycles it would take to severely damage the antibody. It is recommended to aliquot this antibody before freezing. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

Could you give me some information, if any, about cross reactivity studies of this secondary antibody. Thank you very much.

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Answer

Thank you for your enquiry. Mouse IgG antibody (ab6728) has been purified by immunoaffinity chromatography. It has not been preabsorbed. You may wish to take a look at the numerous favourable reviews that we have received about this antibody. You can examine these by clicking on the "Abreviews" tab on the antibody datasheet. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

Dear Karen, Thank you for your reply. I'm thinking of buying the ab18387 and the Mouse IgG antibody (ab6728) as a secondary antibody. They should be fine together in WB analysis. Could you tell me the lifetime of these antibodies?

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Answer

Thank you for your enquiry. Ab18387 is stored at -20C. We generally say that antibodies stored in this way should be good for at least a year. Ab6728 can be stored at either -20C or -80C. The lifetime at -80C would be at least 2 years. Please note that you should aliquot these antibodies before freezing, as repeated freeze-thaw cycles are damaging to the antibody. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Question

May I ask once more? My samples are sheep proteins. I am going to use two antibodies (anti GAPDH and anti GDF8 antibodies). The first antibodies are raised in mouse and rat (mouse IgG and Rat IgG), respectively. There are many secondary antibodies (HRP conjugated) which recognize mouse IgG or rat IgG (ex. ab5870, ab5879, ab5887, ab6728.....). As some of them are raised in goat, I am afraid that I will have a high background. I would like to know which secondary antibodies against mouse and rat IgG is good for western blotting to detect sheep proteins. Thank you for your helps.

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Answer

Thank you for your enquiry. Given that you are using mouse and rat primary antisera and would like to use a secondary antiserum that will not detect any endogenous IgG in your sheep samples, bearing in mind a potential cross reactivity with goat, I would like to recommend the following: ab6728 Mouse IgG antibody (ab6728) HRP conjugated Rabbit ab6820 Mouse IgG antibody (ab6820) HRP conjugated Donkey Both antisera have been raised in rabbit or donkey and are highly unlikely to cross react against sheep. We have also received favourable reviews for them both.

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21-30 of 39 Abreviews or Q&A

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