Recombinant Anti-Wnt3a antibody [EPR21889] (ab219412)

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Western blot - 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: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
Reacts with: Mouse, Rat, Human

Alexa Fluor® 647 APC Carrier Free HRP PE

Product name

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

Rabbit monoclonal [EPR21889] to Wnt3a
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
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 Cyt (intra): 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

Alternative Versions
- Anti-Wnt3a antibody [EPR21889] - BSA and Azide free (ab234099)
- Alexa Fluor® 647 Anti-Wnt3a antibody [EPR21889] (ab270093)
Compatible Secondaries
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Immunohistochemistry reagents
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant mouse Wnt3a protein (ab81484)
Related Products
- Anti-Wnt3 and Wnt3a antibody [EPR4920(2)] (ab172612)
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219412 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/500.
WB		1/1000. Predicted molecular weight: 39 kDa.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/100.
IP		1/30.

Notes
Flow Cyt (Intra) 1/500.
WB 1/1000. Predicted molecular weight: 39 kDa.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/100.
IP 1/30.

Function

Ligand for members of the frizzled family of seven transmembrane receptors. Wnt-3 and Wnt-3a play distinct roles in cell-cell signaling during morphogenesis of the developing neural tube.
Tissue specificity

Moderately expressed in placenta and at low levels in adult lung, spleen, and prostate.
Sequence similarities

Belongs to the Wnt family.
Cellular localization

Secreted > extracellular space > extracellular matrix.
Target information above from: UniProt accession P56704 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 89780 Human
- Entrez Gene: 22416 Mouse
- Entrez Gene: 303181 Rat
- Omim: 606359 Human
- SwissProt: P56704 Human
- SwissProt: P27467 Mouse
- Unigene: 336930 Human
- Unigene: 1367 Mouse
Alternative names
- Protein Wnt 3a Precursor antibody
- Protein Wnt-3a antibody
- Wingless type MMTV integration site family member 3A antibody
- Wnt 3a antibody
- wnt3a antibody
- WNT3A protein antibody
- WNT3A_HUMAN antibody
see all

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 (Intracellular) - Anti-Wnt3a antibody [EPR21889] (ab219412)

Intracellular 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)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab219412? Please let us know so that we can cite the reference in this datasheet.

ab219412 has been referenced in 20 publications.

Yang X et al. WNT/ß-catenin-suppressed FTO expression increases m6A of c-Myc mRNA to promote tumor cell glycolysis and tumorigenesis. Cell Death Dis 12:462 (2021). PubMed: 33966037
Shen X et al. lncRNA NEAT1 facilitates the progression of colorectal cancer via the KDM5A/Cul4A and Wnt signaling pathway. Int J Oncol 59:N/A (2021). PubMed: 34109988
Liu C et al. Tumor-associated macrophage-derived transforming growth factor-ß promotes colorectal cancer progression through HIF1-TRIB3 signaling. Cancer Sci 112:4198-4207 (2021). PubMed: 34375482
Lu X et al. Modulation of HOXA9 after skeletal muscle denervation and reinnervation. Am J Physiol Cell Physiol 318:C1154-C1165 (2020). PubMed: 32233950
Chen H et al. GPX8 is transcriptionally regulated by FOXC1 and promotes the growth of gastric cancer cells through activating the Wnt signaling pathway. Cancer Cell Int 20:596 (2020). PubMed: 33317536

View all Publications for this product

Customer reviews and Q&As

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Immunohistochemistry reagents

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (20)

Customer reviews and Q&As