Donkey Anti-Rabbit IgG H&L (HRP) preadsorbed (ab7083)

Submit a review Q&A (3)References (3)

Overview

  • Product name
    Donkey Anti-Rabbit IgG H&L (HRP) preadsorbed
    See all IgG secondary antibodies
  • Host species
    Donkey
  • 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 ab7083 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 - 1/3000.
IHC-P Use at an assay dependent dilution.
WB 1/1000 - 1/3000.
ELISA 1/7500 - 1/30000.
Immunomicroscopy Use at an assay dependent dilution.
Dot blot Use at an assay dependent dilution.
IHC-Fr Use at an assay dependent dilution.

References

This product has been referenced in:
  • Abdulsid A  et al. Heat shock protein 70 expression is spatially distributed in human placenta and selectively upregulated during labor and preeclampsia. PLoS One 8:e54540 (2013). WB . Read more (PubMed: 23382911) »
  • Faulkner S  et al. A comparison of the bovine uterine and plasma proteome using iTRAQ proteomics. Proteomics 12:2014-23 (2012). Read more (PubMed: 22623423) »
See all 3 Publications for this product

Publishing research using ab7083? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Question

Again, thank you so much for the insight. I did the experiment using both adult rat hippocampus and 3-week-old pup liver against B-actin as control. Apparently, both proteins showed a weak bands, the former was expressed at around ˜35kDa and the latter at around ˜25kDa (which reflected the same results from the first experiment). Could it be that this is actually the folate binding protein (FR-alpha)? It would make sense that the adult rat had a higher kDa post glycosylation. Kindly see the attached file and give us your insight. Thank you so much! :)


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Answer

Thank you for contacting us again and sending this new information.

The WB image looks very convincing and proofs that the antibody is actually picking up its target as it should.

Regarding the double band in the pup sample it may be that the antibody is indeed picking up the non-glycosylated form. In addition, the double band may be due to a low glycosylation level or may be the protein chain plus the C-terminal pro-peptide (approx. 27 kDa).

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

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Question

I truly appreciate your immediate reply especially at this time. From your suggestion, I will try to do a positive control run using a female adult hippocampus sample. Will that be okay? Regarding my inquiry, could you enlighten me on the expression at the lower band around ˜10kDa, whether it is a nonspecific band or is just part of my target protein that has undergone post translational modification? I appreciate if you clarify any of these.:)

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Answer

Thank you for your reply.

Unfortunately, it is quite hard to get some information if this protein expressed in the rat hippocampus. However, I checked the SwissProt site for more information regardingthis for the human protein, and it looks like in human it is only expressed in the cerebellum. I would therefore suggest rather to use a whole brain or cerebellum lysate in case the expression pattern is the same as in humans. Kidneys do also show a high expression level in rats according to UniGene.

To be honest, I cannot tell you if the signal at 10 kDa is specific or not. From checking the SwissProt side, I would say it is probably non-specific, but as I cannot find any information there for the rat protein, it is rather a guess than really knowing. In addition, UniGene reports anon-existing expression level in liver.

Sorry to disappoint here that I cannot help to solve the riddle at the moment.

Please let me know if the new samples provide the expected results.

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Question


I recently ordered Anti-Folate binding protein primary antibody (EPR4708) Cat # ab125030 and donkey anti-rabbit IgG H&L (HRP) secondary anitbody Cat # ab7083 to measure the expression of FBP1 (Folate receptor alpha) that has a predicted band size at 30kDa. The primary antibody I used is predicted to work with rat. I am using a 3-week-old Sprague Dawley rat pup liver sample and did the Western Blot experiment according to the Abcam protocols book (First edition). My B-actin turned out with a specific band at around 43kDa, however, for my target protein FR alpha, 2 nonspecific weak bands (4 columns respective for 4 sample groups) were seen at around 25kDa and 11kDa. Kindly checked the attached file and please let me know whether I can consider any of them as specific to my protein of interest. Thank you so much and hoping for your utmost response. Have a good day!



Sincerely,

Kay





Karen Kay Mejos
Dept. of Nutritional Science and Food Management
Ewha Womans University
11-1 Daehyun-dong, Seodaemun-ku
Seoul, 120-750 Korea
Tel: +82-2-3277-4426
Fax: +82-2-3277-2898
Mobile: 010-9073-9216
e-mail: karenmejos@ewhain.net










Abcam Customer Services and Scientific Support Team
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Answer

Thank you for resending the image. Based on your description of the upper band as being ˜ 25kD it seems this could be the correct band, as the size can vary a few kD just based on the gel, buffer, etc. Adding a negative control could help ensure that this is not a non-specific band. Please let me know if you have any further questions.

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