Antibodies, Proteins, Kits and Reagents for Life Science

351 792 ₸ Product size

100 µl 351 792 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2871] to Ret
Suitable for: ELISA, WB, IP, IHC-P, ICC/IF
Reacts with: Mouse, Rat, Human

Product name

Anti-Ret antibody [EPR2871]
See all Ret primary antibodies
Description

Rabbit monoclonal [EPR2871] to Ret
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Mouse Human
IHC-P	Mouse Human
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human Ret aa 1100 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: P07949
(Peptide available as ab219199)
Positive control
- WB: Neuro-2a, SH-SY5Y and TT cell lysates and mouse and rat brain tissue lysates. IHC-P: Human thyroid gland carcinoma, gastric carcinoma and colon tissues; Mouse colon tissue; Human medullary thyroid carcinoma (MTC) samples from GTCAC haplotype carriers & non-carriers. ICC/IF: Neuro-2a and TT cells. IP: Neuro-2a cell lysate.
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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

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

Protein A purified
Clonality

Monoclonal
Clone number

EPR2871
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ret antibody [EPR2871] (ab134100) Image from Ceolin L et al., PLoS One. 2016;11(2):e0147840. Fig 4.; doi: 10.1371/journal.pone.0147840. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Immunostaining of the Ret proto-oncogene in human Medullary thyroid carcinoma (MTC) samples from GTCAC haplotype carriers & non-carriers.

Two representative slices of Ret Immunostaining in a sample carrier S836S/3’UTR (GTCAC haplotype) (left) and non-carrier of this haplotype (right)
Western blot - Anti-Ret antibody [EPR2871] (ab134100)

Anti-Ret antibody [EPR2871] (ab134100) at 1/1000 dilution (purified) + TT (human thyroid carcinoma cell line) whole cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 124 kDa
Observed band size: 155,175 kDa
why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ret antibody [EPR2871] (ab134100)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue labelling Ret with purified ab134100 at a dilution of 1/50. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-Ret antibody [EPR2871] (ab134100)

Immunocytochemistry/Immunofluorescence analysis of Neuro-2a cells labelling Ret with purified ab134100 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000).

Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Western blot - Anti-Ret antibody [EPR2871] (ab134100)

Anti-Ret antibody [EPR2871] (ab134100) at 1/5000 dilution (purified) + SH-SY5Y (human neuroblastoma cell line from bone marrow) whole cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 124 kDa
Observed band size: 155,175 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ret antibody [EPR2871] (ab134100) This image is courtesy of an anonymous Abreview.

Formaldehyde-fixed, paraffin-embedded human colon tissue stained for Ret using ab134100 at 1/80 dilution in immunohistochemical analysis, followed by Goat anti-Rabbit IgG (HRP antibody.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-Ret antibody [EPR2871] (ab134100)

Immunocytochemistry/Immunofluorescence analysis of TT (human thyroid carcinoma epithelial cell) cells labelling Ret with purified ab134100 at a dilution of 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (1/200) was used as a counter stain.

-ve control: Caco-2 (PMID: 10811228) cells stained with primary antibody (1/100) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000).
Western blot - Anti-Ret antibody [EPR2871] (ab134100)

All lanes : Anti-Ret antibody [EPR2871] (ab134100) at 1/1000 dilution (purified)

Lane 1 : Mouse brain tissue
Lane 2 : Rat brain tissue

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 124 kDa
Observed band size: 155,175 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST
ELISA - Anti-Ret antibody [EPR2871] (ab134100)

ELISA antigen dose-response curve using purified ab134100 at 0-1000 ng/ml. Antigen concentration of 1000 ng/mL. An Alkaline-Phosphatase-conjugated goat anti-rabbit IgG (H+L) (1/2500) was used as the secondary antibody.
Western blot - Anti-Ret antibody [EPR2871] (ab134100)

All lanes : Anti-Ret antibody [EPR2871] (ab134100) at 1/10000 dilution (unpurified)

Lane 1 : Neuro-2a (mouse neuroblastoma cell line) cell lysate
Lane 2 : TT (human thyroid carcinoma cell line) cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 124 kDa
Western blot - Anti-Ret antibody [EPR2871] (ab134100)

All lanes : Anti-Ret antibody [EPR2871] (ab134100) at 1/10000 dilution (unpurified)

Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

Predicted band size: 124 kDa
Observed band size: 155 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and Diluting buffer and concentration: 5% NFDM/TBST
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ret antibody [EPR2871] (ab134100)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid gland carcinoma tissue labelling Ret with unpurified ab134100 at a dilution of 1/250.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ret antibody [EPR2871] (ab134100)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse colon tissue labelling Ret with unpurified ab134100 at a dilution of 1/50 followed by HRP-conjugated goat anti-rabbit IgG (H&L) (ab97051, 1/500). Counter stained with hematoxylin.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Western blot - Anti-Ret antibody [EPR2871] (ab134100)

Anti-Ret antibody [EPR2871] (ab134100) at 1/10000 dilution (purified) + Neuro-2a (mouse neuroblastoma cell line) whole cell lysate at 10 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/2000 dilution

Predicted band size: 124 kDa
Observed band size: 155,175 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking and dilution buffer: 5% NFDM /TBST.

We are unsure about the nature of 40kDa to 60kDa bands, they might be the intracellular fragments of Ret.
Immunoprecipitation - Anti-Ret antibody [EPR2871] (ab134100)

ab134100 (purified) at 1/40 immunoprecipitating Ret in Neuro-2a mMouse neuroblastoma cell line) whole cell lysate.

Lane 1 (input): Neuro-2a whole cell lysate (10µg)

Lane 2 (+): ab134100 + Neuro-2a whole cell lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab134100 in Neuro-2a whole cell lysate.

For western blotting, ab134100 was used at 1/1000 followed by a HRP-conjugated anti-rabbit IgG (specific to the non-reduced form of IgG, 1/1500).

Blocking buffer and dilution concentration: 5% NFDM/TBST.

We are unsure about the nature of 40kDa to 60kDa bands, they might be the intracellular fragments of Ret.
Anti-Ret antibody [EPR2871] (ab134100)

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