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

1-10 of 11 Q&A

Question
Answer

Antibody ab97040 goes through an extra purification step to remove reactivity with immunoglobulins of other species. On the datasheets you will see the following notes in the Purification field.

ab97023 - This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Horse Radish Peroxidase (HRP).

ab97040 - 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).

Read More

Answer

Thank you for contacting us.

Fortunately we offer various protocols and tips that can be found on our webpage under scientific support protocols.

I will give you some links that might be useful for your experiments. However, please note that every antibody and every experiment needs some optimisation. The protocols are just general guidelines.


1. https://docs.abcam.com/pdf/protocols/Permeabilization-and-fixation.pdf

2. https://www.abcam.com/ps/pdf/protocols/ihc_p.pdf (for your questions 1-5)

3. https://www.abcam.com/ps/pdf/protocols/immunostaining_at_a_glance.pdf

4. https://www.abcam.com/ps/pdf/protocols/abcam_troubleshooting_tips_IHC.pdf


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


Free RabMab with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

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Question
Answer

Thank you for confirming that information. I have organised for ab98790 to be sent out to you (Order number xxxxx, Purchase order numberFOCR xxxxx). As we have the Easter bank holidays this will unfortunately only reach you on Wednesday next week.

Hopefully, this new secondary antibody will help the situation. However, if you find the same all over staining please do let me know and we'll take it one reagent at a time to isolate the step which is causing the problems.

I look forward to hearing how you get on.

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Answer

Thank you for getting back to me.

I can arrange for a different secondary antibody to be sent to you. In order to organise this I will need the order numbers of ab78021 and ab97040, or the approximate delivery date and delivery address.

Once I have this information I will look into an appropriate alternative to try.

I look forward to receiving your reply.

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Answer

Thank you for your enquiry.

Please find attached the MSDS for ab97040, which can also be downloaded from the datasheet of this antibody.

Some of our antibodies are produced in-house and others are sourced from suppliers or academic collaborators whose products we believe will be of interest to our customers. All these products are guaranteed by our Abpromise to be of high quality. Our Abpromise assures you that if our products do not perform as our datasheets say they should we offer a replacement or refund within six months of purchase. Some of our collaborators are not certified ISO9001 and so we are not able to provide a certificate of analysis for these particular products.

As the identity of our collaborators is confidential, we provide a certificate of compliance for all of our products available in our catalogue. This document certifies that our products meet the required specifications for a particular lot/batch of product.

The Customer Services and Scientific Support Teams would be very pleased to send a CoC on demand. Could you please let me know if you would like to receive a CoC? If so, could you please confirm the lot number? Unfortunately, x is not a valid lot number for ab97040.

Please do not hesitate to contact me again if you have further questions.

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Answer

Again I apologize for this. As you know these replacement vials are typically sent off very quickly. I have requested (again) that this replacement vial be shipped immediately. I have just received a reply from our UK office stating they are now aware that no replacement has been sent yet. It seems my two inital requests somehow slipped through the cracks. I am optimistic that this time the vial of ab7061 will be sent in the next shipment to Interlab. I am very sorry for this extended delay to you and your customer. I will update you with any further details provided to me. Thanks so much for your patience!

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Answer

Thanks for your emai. I am sorry you have not received the shipment yet.  I requested a replacement of ab7061 be sent on Dec.1. I am looking into this issue and will let you know when to expect this antibody. Thanks for your patience.

Read More

Answer

I have requested that a vial of ab7061 be included in the next shipment to your company for this customer. I hope this is helpful. Please contact me again if you have any further questions.

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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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1-10 of 11 Q&A

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