Antibodies, Proteins, Kits and Reagents for Life Science

526 012 ₸ Product size

200 µg 526 012 ₸ 1 mg 4 690 ₸

Order now and get it on Wednesday March 10, 2021

Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Product name

Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free)
Tested applications

Suitable for: Functional Studies, IHC-P, Flow Cyt, ICC/IF, 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
ab199376 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

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.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C. Do Not Freeze.
Storage buffer

pH: 7.20
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Purity

Protein A purified
Purification notes

Endotoxin level:
Clonality

Monoclonal
Clone number

EPR25A
Isotype

IgG

Immunocytochemistry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Clone EPR25A (ab199376) has been successfully conjugated by Abcam. This image was generated using Rabbit IgG, monoclonal [EPR25A] - Isotype Control (PE). Please refer to ab209478 for protocol details.
Immunofluorescent analysis of HeLa (human cervical cancer) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab209478 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195884, Rat monoclonal [YOL1/34] to Tubulin Microtubule Marker (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).
Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling KCTD5 with ab194825 at 1/70 dilution (red) compared with ab199376 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody. ab199376 detects no signal in FC.
Note: ab199376 detects no signal in Flow cytometry.
Immunocytochemistry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Clone EPR25A (ab199376) has been successfully conjugated by Abcam. This image was generated using Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 647). Please refer to ab199093 for protocol details.
Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199093 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 488), at 1/250 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).
Immunocytochemistry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Clone EPR25A (ab199376) has been successfully conjugated by Abcam. This image was generated using Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Alexa Fluor® 488). Please refer to ab199091 for protocol details.
Immunofluorescent analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cells, fixed with 4% formaldehyde (10 min). The cells were permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199091 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control) at 1/500 dilution (showing no signal) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor^® 594), at 1/250 dilution (pseudocolored in red). Nuclear DNA was labeled with DAPI (shown in blue).

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 100% methanol (5min).
Immunocytochemistry/ Immunofluorescence - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeled with ab199376 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution. Confocal image showing no staining on HeLa cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).
The negative controls are as follows:-
-ve control 1: ab199376 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.
Immunohistochemistry paraffin embedded sections - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Immunohistochemistry analysis of rat colon tissue control for testing ab199376 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody. Counterstained with hematoxylin. No staining on rat colon tissue was observed.
Immunohistochemistry paraffin embedded sections - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Immunohistochemistry analysis of mouse colon tissue control for testing ab199376 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody. Counterstained with hematoxylin. No staining on mouse colon tissue was observed.
Immunohistochemistry paraffin embedded sections - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)

Immunohistochemistry analysis of human colon tissue control for testing ab199376 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. An anti-rabbit IgG H&L (HRP) (ab97051) was used as a secondary antibody. Counterstained with hematoxylin. No staining on human colon tissue was observed.
Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Low endotoxin, Azide free) (ab199376)