Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)

Reviews (2)Q&A (4)References (36)

Overview

  • Product name
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed
    See all IgG secondary antibodies
  • Host species
    Goat
  • Target species
    Rabbit
  • Tested applications
    Suitable for: ICC/IF, IHC-P, WB, ELISA, Immunomicroscopy, Dot blot, IHC-Frmore details
  • Minimal
    cross-reactivity

    Chicken, Cow, Goat, Guinea pig, Hamster, Horse, Human, Mouse, Rat, Sheep more details
  • Immunogen

    Full length native Rabbit IgG (purified).

  • Conjugation
    HRP

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C.
  • Storage buffer
    pH: 7.20
    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 Rabbit IgG coupled to agarose beads.
  • Conjugation notes
    Horseradish Peroxidase (HRP)
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab7090 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/IF 1/1000 - 1/5000.
IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration.
ELISA 1/20000 - 1/80000.
Immunomicroscopy Use at an assay dependent concentration.
Dot blot Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Images

  • Immunohistochemistry - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)
    Immunohistochemistry - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)Lean QY et al. PLoS One (2015)

    Immunohistochemical analysis of healthy and colitis mouse colon sections (untreated and treated with enoxaparin) labeling claudin-4 with ab15104 at 1/200. ab7090, anti-rabbit immunoglobulin G conjugated to horseradish peroxidase (HRP) at 1/300 was used as the secondary antibody. ab64238 at 1/50 was used to develop the histological signal.

    Antigen retrieval was performed by incubating the sections for 10 minutes at 97°C in 1 mM EDTA buffer, pH 8.0 or 10 mM citrate buffer, pH 6.0.

    Control, C; untreated colitis, DSS; oral enoxaparin, OE; intraperitoneal injection of enoxaparin, IPE.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7090)This image is courtesy of an anonymous Abreview.

    ab9986 staining osteoprotegerin in human skin tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% BSA and 5% normal goat serum in PBS for 30 minutes at 22°C; Samples were incubated with primary antibody (1/20) for 9 hours at 4°C. An HRP-conjugated goat anti-rabbit IgG H&L (ab7090) (1/300) was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Chen C  et al. Knockdown of EIF5A2 inhibits the malignant potential of non-small cell lung cancer cells. Oncol Lett 15:4541-4549 (2018). Read more (PubMed: 29541224) »
  • Li Z  et al. Intramedullary central neurocytoma of the thoracic spinal cord: A case report and literature review. Mol Clin Oncol 8:544-548 (2018). Read more (PubMed: 29541463) »
See all 36 Publications for this product

Publishing research using ab7090? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-6 of 6 Abreviews or Q&A

Good abtibody

 Excellent
Abreviews
abreview image
Application
Western blot
I use this antibody mostly in WB. I can get good signals (1/10000 dilution).

Upload image condition:
1st antibody ab108296 1/1000 dilution(5% milk)
2nd antibody This antibody 1/10000 dilution(5% milk)

Abcam user community

Verified customer

Submitted Sep 14 2017

Immunochemistry with ab7090

 Good
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
This is an example of immunochemistry anti-Inos (ab15323) with Goat anti Rabbit HRP (on human artery)
-Deparaffinize
-heat in citrate buffer 20min at 96°C
-H2O2 for 10min at room temperature
-Block with PBS BSA 1% goat serum 5% at least 1h at room temperature
-incubate anti INOS at 1/100 in PBS BSA Overnight at 4°C
-incubate with goat anti rabbit HRP (1/300) for 45min at 37°C
-incubate slides in DAB or other peroxidase substrate
-counterstain with hematoxylin, dehydrate and mount

Abcam user community

Verified customer

Submitted Jan 29 2014

Question

Dear abcam technical support team: This customer already purchases ab80436 (EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit) and she wants to buy the “Goat anti-rabbit HRP Conjugate” alone, could you please offer any suitable selection to her? Thanks for your kindly help

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Answer

Thank you for contacting us.

The Goat anti-rabbit HPR Conjugated used in AB80436 has been developed specifically for use in this kit and is currently not available for separate purchase. We do offer a number of Goat anti-rabbit secondaries which are HRP conjugated and have been validated for used in IHC. AB7090 and AB98512 would each be excellent choices for any IHC against an antibody raised in rabbit. Each are pre-absorbed to reduce cross reactivity, and AB98512 is an F(ab')2 fragment. Use of F(ab')2 fragments further reduce non-specific background as these antibodies lack the Fc region which may bind endogenous Fc receptors within the tissue.

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

I have addressed your questions below:

1. The details you provide indicate several secondaries are giving a band at 75 kDa in the no primary control tests. Please confirm further details of any other secondary antibodies you are having this difficulty with.

a. We have tried secondary antibodies from other vendors (Thermo scientific and from Santa Cruz Biotech) and we get this same band.

I am concerned that something in the sample is crossreacting with the secondary antibodies:

2. Please confirm more details of how the samples are being prepared?

a. The samples are homogenized in RIPA buffer containing a 1% protease and phosphatase inhibitor cocktail (Sigma). Tissue homogenate was centrifuged (16,000g x15 min), and the aqueous layer was transferred to a separate tube and stored at -80°C for later analyses. Thirty μg of total protein was then resolved in a 10% Tris-HCl SDS gel. Proteins were electroblotted using the Bio-Rad Trans Blot SD semi-dry cell onto 0.45 μm nitrocellulose membranes. Membranes were blocked for one hour with one of the following solutions: 3% BSA, 3% dry milk, pierce starting block T20 blocking buffer, pierce protein-free T20 blocking buffer. After the blocking membranes were washed 6 times for 5 minutes, then the membrane was incubated with HRP-conjugated secondary antibody for one hour, re-washed 4 times for 5 minutes, and developed by using the super signal west pico chemiluminescent substrate (Thermo).

3. Is there anything special about these samples? Where the rats treated in any way? Or are the samples from just ordinary rat kidney from an untreated rat?

a. The samples are from two rat strains (insulin resistant rats and their control strain). The insulin resistant rats were supplemented with glucose in their drinking water and treated with an angiotensin receptor blocker.

4. Have any other samples been tried, from a different species or tissue? Does this give the same result?

a. Yesterday I tried a test with kidney and heart samples from this rats and muscle and adipose from elephant seals and the band at 75 kDa appears only in the rat kidney samples. I have attached the image.

5. Have the membranes been stripped and reprobed at all? Or are they fresh membranes?

a. The membranes are fresh.

I look forward to hearing from you.

Thank you,

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Answer

Thank you for your reply.

Reviewing the details, if you obtaining a band at 75 kDa in the no primary control with many secondary antibodies with the rat kidney lysate, but not the other samples, then I would recommend there is something in the rat kidney lysate that is crossreacting with these secondary antibodies.

It would be difficult to determine what the difference with the rat samples may be. However, one suggestion is thatendogenous rat antibody in the sample may be crossreacting with the secondary. Particularly in the case of infection or kidney disease, antibody could build up in the kidney tissue.

In this case:

1. The IgG antibody isotype has a molecular weight of 150 kDa which in reducing and denaturing conditions would break down to heavy and light chains and show on a blot at 25 kDa and 50Kda. However, if not fully reduced anddenatured,the heavy and light chain togethercould possiblyshow at 75 kDa. This may be what you are observing. I can suggest to ensure the sample is fully reduced and denatured when running on the gel.

2. I would suggest to try a pre or cross absorbed antibody to reduce the risk of crossreactivity with the rat IgG in the sample, but it seems you have already done so.

3. In case of problems with the individual rat, or possible contamination of the sample, could you confirm if you have tried running kidney samples from different rats?

I am very sorry there are no further explanations to provide in this case. Regrettably, it seems to be a samplephenomenon with the rat kidneyin this case.

If you have any further questions or concerns, please do not hesitate to contact me and I will be pleased to help.

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Question

Hello technical support,

Recently we purchased two secondary antibodies from abcam to use for western blotting and we are getting a dark band at 75 KD with secondary antibody alone. Can you please advise me on what I should do next.

I have tried many things from trying different blocking solutions to different secondary antibodies, yet I still get the same band. Attached you will find a picture of my results using your secondary antibodies:

On the left is the Mouse monoclonal secondary antibody to rabbit igG light chain (ab99697), and on the right is goat anti-rabbit IgG (HRP) secondary antibody (ab7090).

This is the protocol that I use for the picture that I attached:

Thirty mg of frozen rat kidney tissue was homogenized in 250 μL of RIPA buffer containing a 1% protease and phosphatase inhibitor cocktail (Sigma). Tissue homogenate was centrifuged (16,000g x15 min), and the aqueous layer was transferred to a separate tube and stored at -80°C for later analyses.

Thirty μg of total protein were resolved in a 10% Tris-HCl SDS gel.

Proteins were electroblotted using the Bio-Rad Trans Blot SD semi-dry cell onto 0.45 μm nitrocellulose membranes.

Membranes were blocked with 3% bovine serum albumin (BSA) in PBS containing 0.05% of Tween 20 (PBS-T).

I look forward to hearing from you.

Thank you,

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Answer

Thank you for taking the time to contact us. I am sorry to hear you have had difficulty obtaining satisfactory results from these two secondaryantibody.

The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

I would like to reassure you that both these antibodies aretested and covered by our 6 month guarantee for use in WB. In the event that a product is not functioning in the applications cited on the product data sheet, we will be pleased to provide a credit note or free of charge replacement.

Reviewing this case, I would appreciate if you can confirm some further details. It is very unusual to see this type of problem from several secondary antibodies. Please confirm:

1. The details you provideindicate several secondaries are giving a band at 75 kDa in the no primary control tests. Please confirm further details of any other secondary antibodies you are having this difficultywith.

I am concerned that something in the sample is crossreacting with the secondary antibodies:

2. Please confirm more details of how the samples are being prepared?

3. Is there anything special about these samples? Where the rats treated in any way? Or are the samples from just ordinary rat kidney from an untreated rat?

4. Have any other samples been tried, from a different species or tissue? Does this give the same result?

5. Have the membranes been stripped and reprobed at all? Or are they fresh membranes?

I hope this information is helpful, thank you for your cooperation. I look forward to receiving the requested details which will help us to investigate. I hope we can resolve this case as quickly as possible for you.

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Question

Mr I have some questions about antibodie IgG-ab7090. This antibodie no cross-reactive with mouse monoclonal antibodies (primary antibodies)? How is the difference with secondary rabbit antibodies no pre-adsorbed? I need in my group a secondary rabbit antibodie with not cross reactive with the mouse primary antibodie, especially in inmunoprecipitations protocols for detect in western blot. I need send me this information quickly. Thank you very much

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Answer

This antibody has been pre-adsorbed against Bovine, Chicken, Goat, Guinea Pig, Horse, Human, Mouse, Rat and Sheep Serum Proteins. This minimises the cross reactivity of this antibody with these proteins. However, minimal cross reactivity will still be seen. The non-adsorbed version of this antibody will therefore exhibit a greater cross reactivity to the above proteins than the pre-adsorbed. If you require an antibody towards rabbit IgG that does not react with mouse IgG, then we suggest a "Mouse polyclonal to rabbit IgG", as antibodies from the same species can't react with each other. Unfortunately, we don't stock this product at present.

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