Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20164-278] to Anterior Gradient 2
Suitable for: IHC-P, WB, ICC/IF, Flow Cyt
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-Anterior Gradient 2 antibody [EPR20164-278]
See all Anterior Gradient 2 primary antibodies
Description

Rabbit monoclonal [EPR20164-278] to Anterior Gradient 2
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Mouse Rat Human
WB	Mouse Rat Human Recombinant fragment

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human AGR2 recombinant protein; MCF7 and HT-29 whole cell lysates; Mouse, rat and human stomach lysates; Rat small intestine lysate. IHC-P: Human colon and ovary cancer tissues; Mouse and rat stomach tissues. ICC/IF: MCF7 cells. Flow Cyt: MCF7 cells.
General notes
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.

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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20164-278
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224)

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] (ab209224)

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?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: 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] (ab209224)

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

Blocking/Dilution buffer: 5% NFDM/TBST.

Negative control: HeLa (PMID: 16551856).
Western blot - Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224)

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?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-3: 3 minutes; Lane 4: 10 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224)

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] (ab209224)

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] (ab209224)

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] (ab209224)

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/ Immunofluorescence - Anti-Anterior Gradient 2 antibody [EPR20164-278] (ab209224)

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] (ab209224)

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] (ab209224)

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] (ab209224)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com