Overview

  • Product name

    Goat Anti-Rat IgG H&L (HRP)
    See all IgG secondary antibodies
  • Host species

    Goat
  • Target species

    Rat
  • Specificity

    By immunoelectrophoresis and ELISA this antibody reacts specifically with Rat IgG and with light chains common to other Rat immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins.
  • Tested applications

    Suitable for: ICC, IHC-P, ELISA, WBmore details
  • 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

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

    Molar enzyme/ antibody protein ratio is 4:1
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • General notes

    Part of the AbExcel range.
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab97057 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
ICC Use at an assay dependent dilution.
IHC-P 1/200 - 1/5000.
ELISA 1/10000 - 1/100000. (as primary)
WB 1/2000 - 1/20000. Colorimetric: Use 1/5000-1/30000, Chemiluminescent: use 1/10000-1/50000

Images

  • IHC image of alpha tubulin 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 ab64332, 5µg/ml overnight at +4°C. An HRP-conjugated secondary (ab97057, 1/200 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

  • All lanes : Anti-Tubulin antibody [YL1/2] - Loading Control (ab6160) at 1 µg/ml

    All lanes : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rat IgG H&L (HRP) (ab97057) at 1/2000 dilution
    Lanes 3-4 : Goat Anti-Rat IgG H&L (HRP) (ab97057) at 1/10000 dilution
    Lanes 5-6 : Goat Anti-Rat IgG H&L (HRP) (ab97057) at 1/20000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Observed band size: 52 kDa
    why is the actual band size different from the predicted?



     

    Additional bands at : 85 kDa. We are unsure as to the identity of these extra bands.

References

This product has been referenced in:

  • Tabet R  et al. CUG initiation and frameshifting enable production of dipeptide repeat proteins from ALS/FTD C9ORF72 transcripts. Nat Commun 9:152 (2018). Read more (PubMed: 29323119) »
  • Zhu S  et al. Tanshinone-IIA attenuates the deleterious effects of oxidative stress in osteoporosis through the NF-?B signaling pathway. Mol Med Rep 17:6969-6976 (2018). Read more (PubMed: 29568934) »
See all 30 Publications for this product

Customer reviews and Q&As

1-10 of 15 Abreviews or Q&A

Abreviews
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
The secondary ab works well. Based on my experience for IHC the dilution factor requires optimization based on the tissue and antigen retrieval method used.
In the attached picture you can see F480 used as primary antibody and the secondary Goat Anti-Rat IgG H&L ab 97057 used at the concentration of 1:200.
The dilution 1:500 works well too.

Dr. Francesco Elia Marino

Verified customer

Submitted Apr 15 2016

Abreviews
Application
Western blot
The price of this antibody is good based on the amount provided with the shipment.
I used this Antibody at the dilution 1:10000 and it worked fine in my WB.
In the attached picture ab6640 was used as primary antbody in tissue lysates from human macrophages and ab 97057 was used as secondary antibody. The secondary and primary antibodies were diluted in 2% Milk in PBS.

Dr. Francesco Elia Marino

Verified customer

Submitted Apr 15 2016

Abreviews
Application
Western blot
We are using an 2 years old vial of this antibody, it still works well.

Abcam user community

Verified customer

Submitted Nov 05 2015

Abreviews
Application
Western blot
Antibody recognizes the Rat antibody specifically. I analyzed an HA tagged protein (0.4 OD cells equivalent) and used the anti-Rat HA antibody that was purchased from Roche and this secondary antibody ab97057 was used as secondary. I used 1:10K dilution of the antibody and incubated for 1hr at room temperature. Nitrocellulose membrane was used for blotting. On the blot the lane 1, 2, 3 were three individual clones picked after transformation. The protein of the right molecular weight was observed. (70kD).

Abcam user community

Verified customer

Submitted Mar 26 2014

Abreviews
Application
Western blot
Lane 1 is wild-type mouse ES cells expressing E-cadherin. Lane 2 is an Ecad-/- mES cell line. DECMA-1 primary antibody used (not Abcam). Secondary used at 1:5000 for 1 hour. Use of secondary at higher concentration of 1:2000 results in background staining.

Abcam user community

Verified customer

Submitted Nov 28 2013

Answer

Thank you for your reply. I am sorry that these antibodies did not perform as stated on the datasheet. It is regrettable these antibodies have not worked even though replacements have been tried.

As requested, I have asked our Finance department to issue a credit note for you to the value of one vial ab56299 of and one vial of ab97057 .

Credit note ID: ab23732

The credit note may be used in one of the following ways:

(1) Redeemed against the original invoice if this hasn't already been paid.
(2) Held on the account for use against a future order.
(3) A full refund can be offered where no other invoices are outstanding.

Please contact your Finance department to confirm how you would like the credit note to be used and ensure it is not redeemed without your knowledge.

To specifically receive a refund please ask your Finance department to contact our Finance department at creditcontrol@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website.

The credit note ID is for your reference only, please refer to the credit note ID in any correspondence with our accounting department. We will send you the completed credit note by email or postal mail with the actual credit note number which will start with the letters CGB.

I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service should you require further expert advice

Read More

Answer

Thank you for your message.

I am sincerely sorry to hear this is still not working for you.

I am pleased to raise a credit note for the two items originally purchased. This can be used to pay towards the purchase of any other items from the catalog.

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for confirming these details and for your cooperation.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge alternative replacement of ab98425 with the order number 1180015.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question
Answer

Thank you for your reply.

Any anti rat IgG antibody tested in IHC-P should be suitable for your experiments and would be covered by the guarantee. I can suggest you may like to try the following. I can recommend to review the online datasheet for further information.

ab98425 Goat polyclonal Secondary Antibody to Rat IgG - H+L (HRP), pre-adsorbed
HRP conjugated
Tested in: ELISA, ICC, IHC-P, WB
https://www.abcam.com/index.html?datasheet=98425 (or use the following: https://www.abcam.com/index.html?datasheet=98425).

I hope this will be helpful. I look forward to hearing from you with details of how you would like to proceed.

Read More

Answer

Thank you for your message.

It is indeed disappointing that the results are not yet satisfactory, it is regrettably and I appreciate your concerns.

In this case, I would suggest it would be most beneficial to try a different secondary for detection of the rat IgG2a primary. For example, the following page provides the search results for HRP conjguated anti rat IgG antibodies tested in IHC-P:

https://www.abcam.com/index.html?pageconfig=searchresults

The reason is that by using both a secondary and tertiary antibody, there are more variables added into the experiment which means there is more opportunity for it to go wrong, and it will be much more difficult to troubleshoot. It is also more expensive.

The decision is yours. I am happy to provide a free of charge vial of an HRP conjugated anti rat IgG secondary, and this is what I would advise at this stage. Alternatively, I can provide a free of charge alternative replacement of ab6703. I would like to reassure you that any replacement provided would be covered by the 6 month guarantee.

I hope this will be helpful to you. I look forward to hearing from you with details of how you would like to proceed.

Read More

1-10 of 15 Abreviews or Q&A

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