Anti-SMAD2 Antibody Picoband™ (monoclonal, 3C4)

Product Name

Anti-SMAD2 Antibody Picoband™ (monoclonal, 3C4)

View all Smad2 Antibodies

SKU/Catalog Number

M00090-3

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-SMAD2 Antibody Picoband™ (monoclonal, 3C4) catalog # M00090-3. Tested in Flow Cytometry, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.

Cite This Product

Anti-SMAD2 Antibody Picoband™ (monoclonal, 3C4) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00090-3)

Host

Mouse

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Monoclonal

Clone Number

3C4

Isotype

Mouse IgG1

Immunogen

E.coli-derived human SMAD2 recombinant protein (Position: E83-Q264).

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M00090-3 is reactive to SMAD2 in Human

Applications

M00090-3 is guaranteed for Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

55 kDa

Calculated molecular weight

Background of Smad2

Smad2 (Mothers against decapentaplegic homolog 2), also known as MADR2, MADH2, SMAD family member 2 or SMAD2, is a protein that in humans is encoded by the SMAD2 gene. MAD homolog 2 belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. Eppert et al. mapped the MADR2 gene close to DPC4 at 18q21, a region which is frequently deleted in colorectal cancers. Riggins et al. mapped the human MADH2 gene to 18q21. Nakao et al. refined the localization of the SMAD2 gene to 18q21.1, approximately 3 Mb proximal to DPC4, by fluorescence in situ hybridization. SMAD2 mediates the signal of the transforming growth factor (TGF)-beta, and thus regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. This protein is recruited to the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation (SARA) protein. In response to TGF-beta signal, this protein is phosphorylated by the TGF-beta receptors.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

View more details

Reconstitution

Add 0.2ml of distilled water will yield a concentration of 500ug/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x10⁶ cells, Human

Validation Images & Assay Conditions

Click image to see more details

Figure 1. Western blot analysis of SMAD2 using anti-SMAD2 antibody (M00090-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30ug of sample under reducing conditions.
Lane 1: human HL-60 whole cell lysates,
Lane 2: human Hela whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-SMAD2 antigen affinity purified monoclonal antibody (Catalog # M00090-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for SMAD2 at approximately 55KD. The expected band size for SMAD2 is at 55KD.

Click image to see more details

Figure 2. IHC analysis of SMAD2 using anti-SMAD2 antibody (M00090-3).
SMAD2 was detected in paraffin-embedded section of human gallbladder adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-SMAD2 Antibody (M00090-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

Click image to see more details

Figure 3. IHC analysis of SMAD2 using anti-SMAD2 antibody (M00090-3).
SMAD2 was detected in paraffin-embedded section of human low-differentiated adenocarcinoma of the intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-SMAD2 Antibody (M00090-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

Click image to see more details

Figure 4. IHC analysis of SMAD2 using anti-SMAD2 antibody (M00090-3).
SMAD2 was detected in paraffin-embedded section of human pancreatic cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml mouse anti-SMAD2 Antibody (M00090-3) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.

Click image to see more details

Figure 5. Flow Cytometry analysis of HL-60 cells using anti-SMAD2 antibody (M00090-3).
Overlay histogram showing HL-60 cells stained with M00090-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti- SMAD2 Antibody (M00090-3, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Gene/Protein Information For SMAD2 (Source: Uniprot.org, NCBI)

Alternative Names

hMAD-2; hSMAD2; JV18; JV18-1SMAD, mothers against DPP homolog 2 (Drosophila); MAD homolog 2; Mad protein homolog; Mad, mothers against decapentaplegic homolog 2; MADH2MGC34440; MADR2MGC22139; Mad-related protein 2; mother against DPP homolog 2; mothers against decapentaplegic homolog 2; Mothers against DPP homolog 2; Sma- and Mad-related protein 2; SMAD 2; SMAD family member 2MAD, mothers against decapentaplegic homolog 2 (Drosophila); SMAD, mothers against DPP homolog 2; Smad2; Smad2/3 SMAD2 JV18, JV18-1, MADH2, MADR2, hMAD-2, hSMAD2 SMAD family member 2 mothers against decapentaplegic homolog 2|MAD homolog 2|SMAD, mothers against DPP homolog 2|Sma- and Mad-related protein 2|mother against DPP homolog 2

For more info on SMAD2, check out the SMAD2 Infographic

We have 30,000+ of these available, one for each gene! Check them out.

In this infographic, you will see the following information for SMAD2: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

Take me to the SMAD2 Infographic

Here are featured tools and databases that you might find useful.

• Boster's Pathways Library
• Protein Databases
• Bioscience Research Protocol Resources
• Data Processing & Analysis Software
• Photo Editing Software
• Scientific Literature Resources
• Research Paper Management Tools
• Molecular Biology Software
• Primer Design Tools
• Bioinformatics Tools
• Phylogenetic Tree Analysis