Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free
See all Anterior Gradient 2 primary antibodies
Description

Rabbit monoclonal [EPR20164-278] to Anterior Gradient 2 - BSA and Azide free
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
IHC-P	Human
WB	Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
General notes
Ab251500 is the carrier-free version of ab209224. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with
ab251500 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

Maxpar® is a trademark of Fluidigm Canada Inc.
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: 7.2
Constituent: PBS
Carrier free

Yes
Concentration information loading...
Clonality

Monoclonal
Clone number

EPR20164-278
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human colon is observed [PMID: 24794000]. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

All lanes : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/2000 dilution

Lane 1 : Human AGR2 recombinant protein
Lane 2 : Human AGR3 recombinant protein

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 19 kDa
Observed band size: 17 kDa
why is the actual band size different from the predicted?

This data was developed using ab209224, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lane 1: 10 seconds; Lane 2: 3 minutes.

Human AGR2 recombinant protein contains aa21-175 with a His-tag, and human AGR3 recombinant protein contains aa21-166 with GST-tag.
Western blot - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

All lanes : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/2000 dilution

Lane 1 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Lane 2 : HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 19 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?

Exposure time: 10 seconds

This data was developed using ab209224, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Negative control: HeLa (PMID: 16551856).
Western blot - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

Lanes 1-3 : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/1000 dilution
Lane 4 : Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224) at 1/2000 dilution

Lane 1 : Mouse stomach lysate
Lane 2 : Rat stomach lysate
Lane 3 : Rat small intestine lysate
Lane 4 : Human stomach lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 19 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?

This data was developed using ab209224, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times: Lanes 1-3: 3 minutes; Lane 4: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded human ovary cancer tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human ovary cancer is observed [PMID: 22752467]. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.
Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse stomach is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat stomach is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human pancreas tissue labeling Anterior Gradient 2 with ab209224 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Negative control: human pancreas [PMID: 20550709]. Counter stained with Hematoxylin. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) and HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Anterior Gradient 2 with ab209224 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF7 cell line. Negative control: HeLa (PMID: 16551856). The nuclear counterstain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde-fixed MCF7 (Human breast adenocarcinoma cell line) cells labeling Anterior Gradient 2 with ab209224 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Flow Cytometry - Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)

This data was developed using ab209224, the same antibody clone in a different buffer formulation.Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Anterior Gradient 2 with ab209224 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody. Negative control: HeLa
Anti-Anterior Gradient 2 antibody [EPR20164-278] - BSA and Azide free (ab251500)