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  1. Link

    rabbit-mouse-igg-hl-hrp-ab6728.pdf

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Immunology Immunoglobulins Heavy Chain IgG
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Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

  • Datasheet
  • SDS
Reviews (10)Q&A (31)References (779)

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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

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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

    • Immunology
    • Immunoglobulins
    • Heavy Chain
    • IgG
    • Secondary antibodies
    • anti-Mouse
    • IgG
    • Enzyme
    • HRP

Associated products

  • Substrate reagent

    • TMB ELISA Substrate (Highest Sensitivity) (ab171522)
    • TMB ELISA Substrate (High Sensitivity) (ab171523)
    • TMB ELISA Substrate (Fast Kinetic Rate) (ab171524)
    • TMB ELISA Substrate (Slow Kinetic Rate) (ab171525)
    • TMB ELISA Substrate (Slower Kinetic Rate) (ab171526)
    • TMB ELISA Substrate (Slowest Kinetic Rate) (ab171527)

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 (10)
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

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (779)

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

ab6728 has been referenced in 779 publications.

  • Hu X & Miao H MiR-539-5p inhibits the inflammatory injury in septic H9c2 cells by regulating IRAK3. Mol Biol Rep 49:121-130 (2022). PubMed: 34757596
  • Dou JY  et al. Betulin Targets Lipin1/2-Meidated P2X7 Receptor as a Therapeutic Approach to Attenuate Lipid Accumulation and Metaflammation. Biomol Ther (Seoul) 30:246-256 (2022). PubMed: 34815367
  • Sun H  et al. Long non-coding RNA H19 mediates N-acetyltransferase 1 gene methylation in the development of tamoxifen resistance in breast cancer. Exp Ther Med 23:12 (2022). PubMed: 34815764
  • Liu J  et al. Study on Toll-Like Receptor 2-Mediated Inflammation-Induced Familial Hypertension Combined with Hyperlipemia and Its Mechanism. J Healthc Eng 2022:1473597 (2022). PubMed: 35035808
  • Zhao F  et al. LINC02190 inhibits the embryo-endometrial attachment by decreasing ITGAD expression. Reproduction 163:107-118 (2022). PubMed: 35038314
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

31-40 of 41 Abreviews or Q&A

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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Abcam community

Verified customer

Asked on Jan 09 2006

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.

Read More

Abcam Scientific Support

Answered on Jan 09 2006

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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Abcam community

Verified customer

Asked on Nov 14 2005

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.

Read More

Abcam Scientific Support

Answered on Nov 14 2005

Question

Thank you for reply. I have one more question. I want to use ab6728, Rabit anti mouse IgG + HRP, as a secondary Ab.Do you know if it has been rat absorbed(No cross-reactivity with rat tissue). Best regards,

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Abcam community

Verified customer

Asked on Sep 28 2005

Answer

Thank you for your email. No, ab6728 is not pre-adsorbed. If you have any additional questions, please contact us again.

Read More

Abcam Scientific Support

Answered on Sep 28 2005

Question

I would like to know the availability of ab11026 and ab6728 as well as the current pricing. I would like to use these antibodies for Western blots of E. coli cell lysates that have had the human AQP4 gene inserted, in order to assess expresion levels of hAQP4 (that are currently very low). I would like to achieve the highest sensitivity possible. The hAQP4 proteins are 6-His tagged and I have currently been using antibodies to the His-tag, but have not been able to detect the expression of any hAQP4. I was planning to use your ab11026 as a primary antibody to hAQP4, use the ab6728 (with HPR conjugated)as the secondary antibody, and then use a chemiluminescent reagent for signal. Does this sound like a smart way to go? Any suggestions that you can offer will be greatly appreciated. Thank you.

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Abcam community

Verified customer

Asked on Apr 19 2005

Answer

Thank you for your email. Ab11026 - Mouse monoclonal antibody to Aquaporin 4 - is currently in stock and is $299 for 50 ug. Ab6728 - Rabbit polyclonal antibody to Mouse IgG H&L (HRP) - is also currently in stock and is $150 for 1 mg. Ab11026 has been tested for application in Western blotting and found to cross-react with human. If you go ahead and try this antibody with your samples, I do suggest running a positive control; kidney tissue is recommended. Ab6728 is a suitable secondary antibody to use with this primary. If you have any additional questions, please contact us again.

Read More

Abcam Scientific Support

Answered on Apr 19 2005

Question

1:250 dilution was used and I have done an overnight incubation. Looking forward to hearing from you soon.

Read More

Abcam community

Verified customer

Asked on Mar 26 2005

Answer

Thank you for your email. At this point I would like to offer you a replacement vial free of charge or a refund, please let me know which you would prefer. Thanks again, and I look forward to hearing from you.

Read More

Abcam Scientific Support

Answered on Mar 30 2005

Question

Thank you for the suggestion. After repeating the experiment using HUVEC postive control (loading 20, 50 and 100ug of lysate) yesterday, we still faced the same problem. Only a very very faint band was observed when using 50 and 100 ug HUVEC lysate after 1 hour exposure. Please find below answers in blue. It seems that the human placental growth factor antibody is not performing satisfactory. Please assist us for further troubleshooting. Looking forward to hearing from you soon. Hope you have a great easter holiday.

Read More

Abcam community

Verified customer

Asked on Mar 24 2005

Answer

Thank you for your email. I'm replying to with regard to this issue as my colleague Tanya is away for the Easter holidays. With the HUVEC lysate, what dilutions of ab10966 did you try? Did you incubate overnight at 4C? I apologize if this information was in your previous email, the bottom may have been accidently deleted. Thanks again, and have a great weekend.

Read More

Abcam Scientific Support

Answered on Mar 25 2005

Question

I have purchased 2 antibodies from Abcam but a very very faint band (almost clear film) was observed using AB10966. Kerationcyte lsyate was used and loaded 100ug. Preparation of lysate Remove medium and wash 3x PBS, each wash 5ml. Add in 50ul of lysis buffer (+ Protease inhibitor) to each plate and scrape cells. Collect cell lysate in a tube and syringe it 3x. Spin at 11,000rpm for 5 min to remove cell debris. Collect supernatent and do protein assay. Samples were heat for 5 min together with sample buffer before loading into the gel. Primary Ab used : Ab10966 (lot no 95559) 1: 500 & 1:250 dilutions were performed. Incubate for 1 hr at room temp with shaking, wash 3X 5min with TBS Tween. Secondary Ab Used: Ab6728 Incubate for 1 hr at room temp with shaking, wash 3X 10min with TBS Tween. Detection Method ECL Western Blotting reagent A bark band was seen when using the same secondary antibody for other experiment. No background were seen, even band of intrest is not lit up dark enough when press for 1 hr. 3 times for repeating this experiment - all results obtained are the same (almost clear film obtained). The only differences in this 3 experiment is the different dilution tested for the primary antibody. No positive controls were used, however paper states that keratinocytes will produce PLGF!!

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Abcam community

Verified customer

Asked on Mar 17 2005

Answer

Thank you for contacting us and for providing details of your Western blot protocol. We are very sorry to hear that you are having problem with this antibody. We have read through your message and would like to make the following suggestions/comments: 1. Although we suggest using this antibody at a concentration of 1 - 2 µg/ml, this range is just a recommendation. This product was tested and characterised using recombinant protein. The detection limit for recombinant human placental growth factor is approximately 20 ng/lane and 300 ng/lane under non-reducing and reducing conditions, respectively. We would strongly suggest running purified recombinant protein as positive control along with your samples. 2. If this target protein is expressed at lower levels, then you need to optimize the concentration. Do you know the expressed levels of the Placental Growth Factor in the keratinocytes? If you can't have recombinant protein, then you need to use Human umbilical vein endothelial cells as positive control. 3. You have not mentioned in your e-mail from which species the cells come from? Are they human, mouse, rat etc? This antibody recognizes only human Placental Growth Factor, we have not tested the cross-reactivity with other species. 4. Always use freshly made lysis buffer with a cocktail of proteinase inhibitors. We normally suggest using RIPA buffer. 5. Incubate the membrane with the primary antibody overnight at 4oC to see if the signal is getting stronger or not. 6. Have you tested the transfer of the proteins with Ponceau or other type of dye? How do the molecular weight markers look like? 7. Please also test the secondary antibody with another primary antibody. 8. Finally, we would suggest testing whole cell lysate (make samples before the centrifugation step in case the target you may lose some material in the pellet). Load 20-30 ug per ml of total protein per lane but not more than that other wise the gel may be overloaded. We hope this information will be useful for you. Please find enclose Preparation of RIPA cell lysates for Western Blots: 1. Wash cell pellets once with ice-cold PBS. 2. Add 1 ml of RIPA buffer to 10x8 cells, incubate on ice for 20 min, vortex 2 to 3 times. 3. Centrifuge for 5 min at 4oC at maximum speed in a microfuge tube. 4. Transfer supernatant into clean tube. Measure protein concentration with a protein assay. 5. Adjust concentration to 5 mg/ml with RIPA lysis buffer. 6. Add equal volume of 2 x SDS sample buffer into cell lysate, boil for 5 min. 7. Store at -20oC for daily use or -80oC for long term. Avoid repeated freeze thaw cycles. RIPA Base Ingredients Tris-HCl: 50 mM, pH 7.4 NP-40: 1% Na-deoxycholate: 0.25% NaCl: 150 mM EDTA: 1 mM PMSF: 1 mM Aprotinin, leupeptin, pepstatin: 1 microgram/ml each RIPA Protease Inhibitors Phenylmethylsulfonyl fluoride (PMSF) (200 mM stock solution in isopropanol; store at room temperature) EDTA (calcium chelator; 100 mM stock solution in H2O, pH 7.4) Leupeptin (store frozen in aliquots, 1 mg/ml in H2O) Aprotinin (store frozen in aliquots, 1 mg/ml in H2O) Pepstatin (store frozen in aliquots, 1 mg/ml in methanol) Good luck! Should you need further assistance, then please do not hesitate to contact us again.

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Abcam Scientific Support

Answered on Mar 22 2005

Question

We would be using your priomary for estradiol. Would we need to conjugate some estradiol for this to work?

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Abcam community

Verified customer

Asked on Mar 14 2005

Answer

Thank you for your email. You don't have to conjugate the primary antibody, you can do an indirect ELISA where you use an alkaline phosphatase-conjugated secondary antibody or peroxidase-conjugated secondary antibody. Below is our protocol for indirect ELISA. Please contact us again if you have any additional questions. ELISA 1. Dilute the antigen to a final concentration of 20 µg/ml in PBS. Coat the wells of a PVC microtiter plate with the antigen by pipeting 50 µl of the antigen dilution per well. 2. Cover the plate with an adhesive plastic and incubate for 2 h at room temperature. 3. Remove the coating solution and wash the plate twice by filling the wells with 300 µl PBS. The solutions or washes are removed by flicking the plate over a sink. The remaining drops are removed by patting the plate on a paper towel. 4. Block the remaining protein-binding sites in the coated wells by adding 300 µl blocking buffer, 5% non fat dry milk/PBS, per well. 5. Cover the plate with an adhesive plastic and incubate for at least 2 h at room temperature or, if more convenient, overnight at 4°C. 6. Wash the plate twice with PBS. 7. Make 10-fold dilutions (1:100, 1:1,000, 1:10,000, 1:100,000 and 1:1,000,000) of serum and pre-immune serum (the latter as a negative control) in blocking buffer. Add 50 µl of each dilution to an antigen-coated well. 8. Cover the plate with an adhesive plastic and incubate for 2 h at room temperature. 9. Wash the plate four times with PBS. 10. Add 50 µl of secondary antispecies antibody conjugated to alkaline phosphatase, diluted at the optimal concentration (according to the manufacturer) in blocking buffer immediately before use. 11. Cover the plate with an adhesive plastic and incubate for 2 h at room temperature. 12. Wash the plate four times with PBS. 13. Dissolve p-Nitrophenyl phosphate at a concentration of 1 mg/ml in substrate buffer (1M diethanolamine, 0.5 mM MgCl2, pH 9.8). Add 50 µl of the substrate solution per well with a multichannel pipet or a multipipet. 14. Measure the absorbance at 405 nm, using a microtiter plate spectrophotometer. Perform an end-point measurement after 1 h. 15. Calculate the titer of the sera. The titer can be defined as the dilution of serum giving an optical density (OD) of 0.2 above the background of the ELISA after a 1-h reaction.

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Abcam Scientific Support

Answered on Mar 15 2005

Question

What HRP antibody do you recommend using as the labeled antibody for ELISAs?

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Abcam community

Verified customer

Asked on Mar 14 2005

Answer

Thank you for your enquiry. It depends on what the primary antibody is - would you be using a mouse monoclonal? If you are, this is a good one to try. Just so you know, at the bottom of the datasheets for all our primary antibodies there is a list of compatible secondaries which can be used.

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Abcam Scientific Support

Answered on Mar 14 2005

Question

Please let me know what dilution of ab6728 (anti-mouse secondary) you use for westerns? Thanks,

Read More

Abcam community

Verified customer

Asked on Dec 06 2004

Answer

Thank you for your email. We have received feedback from customers who have used this antibody that dilutions from 1:2000 to 1:5000 have worked very well for them. You may read these reviews by clicking on the reviews tab located on the online datasheet for ab6728. I would therefore suggest starting with a dilution of 1:3000 and optimizing from there. If you have any additional questions, please contact us again.

Read More

Abcam Scientific Support

Answered on Dec 06 2004

31-40 of 41 Abreviews or Q&A

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