Overview

  • Product name
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed
    See all IgG secondary antibodies
  • Description
    Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed
  • Host species
    Goat
  • Target species
    Mouse
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with Mouse IgG and with light chains common to other Mouse immunoglobulins. No antibody was detected against non immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, goat, horse, human, pig, rabbit and rat IgG was detected.

     

  • Tested applications
    Suitable for: IHC-P, ELISA, WBmore details
  • Minimal
    cross-reactivity

    Chicken, Cow, Goat, Horse, Human, Pig, Rabbit, Rat, Sheep more details
  • Immunogen

    Other Immunogen Type corresponding to Mouse IgG.

  • Conjugation
    HRP

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    Preservative: 0.1% Proclin
    Constituents: 0.2% BSA, PBS
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Purification notes
    Antiserum was cross adsorbed using bovine, chicken, horse, human, pig, rabbit and rat immunosorbents to remove cross reactive antibodies. This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Horse Radish Peroxidase (HRP).
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab97040 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
IHC-P 1/200 - 1/5000.
ELISA 1/10000 - 1/100000. (Primary).
WB 1/5000 - 1/50000. For colormetric detection 1/5000 - 1/30000. For chemiluminescent dectection 1/10000 - 1/50000.

Images

  • All lanes : Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : PC12 (Rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 150 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab7291 overnight at 4°C. Antibody binding was detected using an anti-mouse HRP (ab97040), and visualised using ECL development solution ab133406

  • Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed (ab97040) was used at 1/5000 dilution developed using the ECL technique, with ab86741.

  • IHC image of beta actin staining in human colon formalin fixed paraffin embedded tissue section*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8224, 3µg/ml overnight at +4°C. An HRP-conjugated secondary (ab97040, 1/500 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.

    The inset negative control image is taken from an identical assay without primary antibody.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

References

This product has been referenced in:
  • Gao P  et al. SNHG20 serves as a predictor for prognosis and promotes cell growth in oral squamous cell carcinoma. Oncol Lett 17:951-957 (2019). Read more (PubMed: 30655853) »
  • Deng R  et al. lncRNA SNHG1 negatively regulates miRNA-101-3p to enhance the expression of ROCK1 and promote cell proliferation, migration and invasion in osteosarcoma. Int J Mol Med 43:1157-1166 (2019). Read more (PubMed: 30592267) »
See all 60 Publications for this product

Customer reviews and Q&As

11-13 of 13 Abreviews or Q&A

Question
Answer

Thank you for contacting us. Please find attached. 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

LOT NUMBER gr39048-5 ORDER NUMBER 952354 DESCRIPTION OF THE PROBLEM High background and no band The result of using ab97040 as the 2nd Ab presents high background while the other 2nd Ab (AP124P) works well with the same sample and primary antibody. I also tries to lower the concentration of 1st Ab and 2nd Ab, and also block at 4°c O/N although, the background decreases but it is still to high to interfere the target signal. Furthermore, the signal is weaker comparing to the result with the other 2nd Ab (AP124P). SAMPLE • Species: rats (This customer has used the 1st Ab : “Monoclonal • Anti-human Chordin-Like 1 Antibody” to detect expression of protein in Rat sample, and he claims it did work!) • What’s cell line or tissue: ovary tissue lysate • Cell extract or Nuclear extract: cell • Purified protein or Recombinant protein: PRIMARY ANTIBODY R&D mouse ANTI-human chordin-like 1 (cat:MAB1808) http://www.rndsystems.com/pdf/MAB1808.pdf • Species: mouse • Reacts against: human • At what dilution(s) have you tested this antibody: 1:500 • What dilution buffer was used: 5% milk • Incubation time: o/n • Incubation temperature: 4oC • What washing steps were done: PBST 3 times DETECTION METHOD ECl POSITIVE AND NEGATIVE CONTROLS USED Yes, the another 2nd Ab works well with the same sample and 1st Ab. CHEMICON goat anti mouse HRP (cat: AP124P) http://www.millipore.com/catalogue/item/ap124p ANTIBODY STORAGE CONDITIONS 4゚C SAMPLE PREPARATION • What lysis buffer was used: using the PBS and protease inhibitors to extract the total protein from tissue • What protease inhibitors were used: roche complete protease inhibitor cocktail • What loading buffer was used: 4x sample buffer • Phosphatase inhibitors: no • Did you heat the samples: temperature and time: 100oC 5min AMOUNT OF PROTEIN LOADED • How much protein was loaded: 15~90 ug ELECTROPHORESIS/GEL CONDITIONS • Reducing or non reducing gel: Reducing • Reducing agent: SDS • Gel percentage : 12% TRANSFER AND BLOCKING CONDITIONS • Transfer conditions: (Type of membrane, Protein transfer verified): PVDF Blocking conditions • Buffer: PBS • Blocking agent: milk, BSA, serum, what percentage: 5% milk • Incubation time: 1h • Incubation temperature: RT SECONDARY ANTIBODY Secondary Antibody: ab97040 • Species: goat • Reacts against: mouse • At what dilution(s) have you tested this antibody: 1:20000~1:100000 • Incubation time: 1hr , o/n • Wash steps: PBST 3 time • Fluorochrome or enzyme conjugate: HRP • Do you know whether the problems you are experiencing come from the secondary? We have tried our original 2nd antibody for many times, and it does work in our experiment condition. However, your 2nd antibody (ab97040), with different dilution ratio and sample concentration, no specific bands were detected. 2nd Ab: CHEMICON goat anti mouse HRP (cat: AP124P) http://www.millipore.com/catalogue/item/ap124p HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Dilution ratio, dilution buffer, sample concentration.

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Answer

On Monday, November 7 I sent an email offering to replace or credit the purchase of ab97040 as it is not working as expected. When you have a moment, please let me know how you wish to proceed and I will process your request immediately.

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Question

LOT NUMBER gr39048-5 ORDER NUMBER 952354 DESCRIPTION OF THE PROBLEM High background and no band The result of using ab97040 as the 2nd Ab presents high background while the other 2nd Ab (AP124P) works well with the same sample and primary antibody. I also tries to lower the concentration of 1st Ab and 2nd Ab, and also block at 4°c O/N although, the background decreases but it is still to high to interfere the target signal. Furthermore, the signal is weaker comparing to the result with the other 2nd Ab (AP124P). SAMPLE • Species: rats (This customer has used the 1st Ab : “Monoclonal • Anti-human Chordin-Like 1 Antibody” to detect expression of protein in Rat sample, and he claims it did work!) • What’s cell line or tissue: ovary tissue lysate • Cell extract or Nuclear extract: cell • Purified protein or Recombinant protein: PRIMARY ANTIBODY R&D mouse ANTI-human chordin-like 1 (cat:MAB1808) http://www.rndsystems.com/pdf/MAB1808.pdf • Species: mouse • Reacts against: human • At what dilution(s) have you tested this antibody: 1:500 • What dilution buffer was used: 5% milk • Incubation time: o/n • Incubation temperature: 4oC • What washing steps were done: PBST 3 times DETECTION METHOD ECl POSITIVE AND NEGATIVE CONTROLS USED Yes, the another 2nd Ab works well with the same sample and 1st Ab. CHEMICON goat anti mouse HRP (cat: AP124P) http://www.millipore.com/catalogue/item/ap124p ANTIBODY STORAGE CONDITIONS 4゚C SAMPLE PREPARATION • What lysis buffer was used: using the PBS and protease inhibitors to extract the total protein from tissue • What protease inhibitors were used: roche complete protease inhibitor cocktail • What loading buffer was used: 4x sample buffer • Phosphatase inhibitors: no • Did you heat the samples: temperature and time: 100oC 5min AMOUNT OF PROTEIN LOADED • How much protein was loaded: 15~90 ug ELECTROPHORESIS/GEL CONDITIONS • Reducing or non reducing gel: Reducing • Reducing agent: SDS • Gel percentage : 12% TRANSFER AND BLOCKING CONDITIONS • Transfer conditions: (Type of membrane, Protein transfer verified): PVDF Blocking conditions • Buffer: PBS • Blocking agent: milk, BSA, serum, what percentage: 5% milk • Incubation time: 1h • Incubation temperature: RT SECONDARY ANTIBODY Secondary Antibody: ab97040 • Species: goat • Reacts against: mouse • At what dilution(s) have you tested this antibody: 1:20000~1:100000 • Incubation time: 1hr , o/n • Wash steps: PBST 3 time • Fluorochrome or enzyme conjugate: HRP • Do you know whether the problems you are experiencing come from the secondary? We have tried our original 2nd antibody for many times, and it does work in our experiment condition. However, your 2nd antibody (ab97040), with different dilution ratio and sample concentration, no specific bands were detected. 2nd Ab: CHEMICON goat anti mouse HRP (cat: AP124P) http://www.millipore.com/catalogue/item/ap124p HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 5 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? Dilution ratio, dilution buffer, sample concentration.

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Answer

Thank you for submitting your completed questionnaire and for including the western blot images. I agree that the vial of ab97040 you received is giving unreasonably high background. I would be happy to offer a replacement or credit. Please let me know how you wish to proceed.

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11-13 of 13 Abreviews or Q&A

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