Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Wnt3a antibody [EPR21889] (ab219412)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR21889] to Wnt3a
Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Product name

Anti-Wnt3a antibody [EPR21889]
See all Wnt3a primary antibodies
Description

Rabbit monoclonal [EPR21889] to Wnt3a
Host species

Rabbit

Tested Applications & Species

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

See all applications and species data

Immunogen

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: L Wnt-3a (Wnt-3a stably transfected Mouse subcutaneous connective tissue areolar and adipose fibroblast) supernatant and whole cell lysate. His-tagged human Wnt3a recombinant protein (aa19-352). Human fetal brain and fetal skin lysate. HCT 116 whole cell lysate. IHC-P: Mouse and rat E14.5 tissue. ICC/IF: L Wnt-3a cells. Flow Cytometry: L Wnt-3a cells. IP: L Wnt-3a whole 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.

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: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21889
Isotype

IgG
Research areas

Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)

All lanes : Anti-Wnt3a antibody [EPR21889] (ab219412) at 1/1000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : Human fetal skin lysate
Lane 3 : HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate

Lysates/proteins at 10 µg per lane.

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

Predicted band size: 39 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Flow Cytometry - Anti-Wnt3a antibody [EPR21889] (ab219412)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) cell line (right) labeling Wnt3a with ab219412 at 1/500 compared with a Rabbit monoclonal IgG (ab172730) (left). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

Two Wnt3a-expressing populations can be seen labeled in this stably transfected cell line.
Immunoprecipitation - Anti-Wnt3a antibody [EPR21889] (ab219412)

Wnt3a was immunoprecipitated from 0.35 mg L Wnt-3a (Wnt-3a stably-transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) whole cell lysate with ab219412 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219412 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: L Wnt-3a (Wnt-3a stably-transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) whole cell lysate 10 µg (Input).
Lane 2: ab219412 IP in L Wnt-3A whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219412 in L Wnt-3a whole cell lysate.
Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 10 seconds.
Immunocytochemistry/ Immunofluorescence - Anti-Wnt3a antibody [EPR21889] (ab219412)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) cells labeling Wnt3a with ab219412 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in L Wnt-3A cell line. The nuclear counter stain is DAPI (blue).
Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)

Immunohistochemical analysis of paraffin-embedded rat E14.5 tissue labeling Wnt3a with ab219412 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on the hem remnant of rat E14.5 (PMID: 15509764; PMID: 16410414) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3a antibody [EPR21889] (ab219412)

Immunohistochemical analysis of paraffin-embedded mouse E14.5 tissue labeling Wnt3a with ab219412 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on the hem remnant of mouse E14.5 (PMID: 15509764; PMID: 16410414) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)

All lanes : Anti-Wnt3a antibody [EPR21889] (ab219412) at 1/20000 dilution

Lane 1 : His-tagged human Wnt3 recombinant protein (aa22-355), 40 ng
Lane 2 : His-tagged human Wnt3a recombinant protein (aa19-352), 20 ng

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

Predicted band size: 39 kDa

Exposure time: 3 minutes

Blocking/ Dilution buffer and concentration: 5% NFDM/TBST.
Western blot - Anti-Wnt3a antibody [EPR21889] (ab219412)

All lanes : Anti-Wnt3a antibody [EPR21889] (ab219412) at 1/1000 dilution

Lane 1 : L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) supernatant at 20 µl
Lane 2 : L Wnt-3a (Wnt-3a stably transfected mouse subcutaneous connective tissue areolar and adipose fibroblast) whole cell lysate at 20 µg

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

Predicted band size: 39 kDa

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

Exposure times: Lane 1: 3 minutes; Lane 2: 3 seconds.
Anti-Wnt3a antibody [EPR21889] (ab219412)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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