Name: Rabbit IgG, monoclonal [EPR25A] - Isotype Control
Brand: RabMAb
SKU: ab172730

Product name

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

Suitable for: ICC/IF, Flow Cyt, IHC-P, CHIPseq, 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.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab^® patents.
This product is a recombinant rabbit monoclonal antibody.

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

pH: 7.20
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

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. Please note: This product should be diluted to the same concentration (not dilution) of the primary antibody to be used.
CHIPseq		Use at an assay dependent concentration. PubMed: 26455392
IP		Use at an assay dependent concentration.

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.
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).
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).
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 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)

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.
Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunocytochemsitry/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).
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 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.
Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Immunocytochemsitry/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).
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

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols

This product has been referenced in:

Li B et al. Decreased H3K9ac level of AT2R mediates the developmental origin of glomerulosclerosis induced by prenatal dexamethasone exposure in male offspring rats. Toxicology 411:32-42 (2019). Read more (PubMed: 30359671) »
Liang W et al. Knockdown of growth-arrest specific transcript 5 restores oxidized low-density lipoprotein-induced impaired autophagy flux via upregulating miR-26a in human endothelial cells. Eur J Pharmacol 843:154-161 (2019). Read more (PubMed: 30468731) »

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