Rabbit IgG, monoclonal [EPR25A] - Isotype Control Recombinant (ab172730)

Product name

Rabbit IgG, monoclonal [EPR25A] - Isotype Control
Tested applications

Suitable for: ICC/IF, Flow Cyt, IHC-P, ChIP-sequencing, IPmore details
Immunogen

Chemical/ Small Molecule conjugated to keyhole limpet haemocyanin. KLH is a copper containing oxygen carrier occurring freely dissolved in the hemolymph of many molluscs and arthropods. KLH forms a large complex composed of ~50 kDa subunits.
General notes
KLH is often used in molecular immunology as a carrier protein conjugated to low molecular weight molecules such as peptides, amino acids, nucleic acids, drugs or toxins to render them more immunogenic due to the size of the conjugate complex and the immunogenicity of KLH.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR25A
Isotype

IgG

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab172730 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		Use at an assay dependent concentration. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
Flow Cyt		Use at an assay dependent concentration. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
IHC-P		Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
ChIP-sequencing		Use at an assay dependent concentration. PubMed: 26455392
IP		Use at an assay dependent concentration.

Notes
ICC/IF Use at an assay dependent concentration. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
Flow Cyt Use at an assay dependent concentration. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
ChIP-sequencing Use at an assay dependent concentration. PubMed: 26455392
IP Use at an assay dependent concentration.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Vimentin RabMAb (ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).
Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

LINE-1 ORF1p was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with ab216324 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216324 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: F9 whole cell lysate 10 µg (Input).
Lane 2: ab216324 IP in F9 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216324 in F9 whole cell lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 30 seconds.
Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

ab124962 (purified) at 1/20 immunoprecipitating IL-1RA in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 whole cell lysate (10µg)
Lane 2 (+): ab124962 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124962 in NIH/3T3 whole cell lysate.
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab133557 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab133557 at 1/60 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

Conjugated versions are available for this clone: Alexa Fluor^® 488 (ab199091), Alexa Fluor^® 647 (ab199093), R-PE (ab209478), APC (ab232814).
Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Overlay histogram showing K562 (human chronic myelogenous leukemia) cells stained with ab196018 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab196018 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Vimentin RabMAb (ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with unpurified Rabbit IgG ab172730 at 1/10. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with purified Rabbit IgG ab172730 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.
Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Overlay histogram showing SH-SY5Y (human neuroblastoma) cells stained with ab179513 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab179513 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.
Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab185633 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab185633 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter
Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunofluorescent staining of HeLa cells using anti-AIF RabMAb (ab32516, left panel) (green) and Rabbit mAb IgG control (ab172730, right panel). DAPI nuclear staining (blue).

Conjugated versions are available for this clone: Alexa Fluor^® 488 (ab199091), Alexa Fluor^® 647 (ab199093), R-PE (ab209478), APC (ab232814).
Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunocytochemistry/immunofluorescence analysis of HeLa cells with purified Rabbit IgG ab172730 at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).
Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunocytochemistry/immunofluorescence analysis of HeLa cells with unpurified Rabbit IgG ab172730 at 1/10. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).
Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Publishing research using ab172730? Please let us know so that we can cite the reference in this datasheet.

ab172730 has been referenced in 97 publications.

Shu L et al. lncRNA ANRIL protects H9c2 cells against hypoxia-induced injury through targeting the miR-7-5p/SIRT1 axis. J Cell Physiol 235:1175-1183 (2020). PubMed: 31264206
Pillai-Kastoori L et al. Antibody validation for Western blot: By the user, for the user. J Biol Chem 295:926-939 (2020). PubMed: 31819006
Chen Z et al. lncRNA HOTAIRM1 promotes osteogenesis of hDFSCs by epigenetically regulating HOXA2 via DNMT1 in vitro. J Cell Physiol N/A:N/A (2020). PubMed: 32324272
Uysal F & Ozturk S The loss of global DNA methylation due to decreased DNMT expression in the postnatal mouse ovaries may associate with infertility emerging during ovarian aging. Histochem Cell Biol N/A:N/A (2020). PubMed: 32514790
Li L et al. miR-148a/LDLR mediates hypercholesterolemia induced by prenatal dexamethasone exposure in male offspring rats. Toxicol Appl Pharmacol 395:114979 (2020). PubMed: 32234517

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Overview

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Tested applications

Immunogen

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

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References (97)

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