SMAD6 Antibody - BSA Free

Novus Biologicals | Catalog # NB100-56440

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Monkey, Primate, Sheep

Cited:

Human, Mouse, Rat, Ovine

Predicted:

Equine (92%). Backed by our 100% Guarantee.

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Immunohistochemistry-Paraffin, Western Blot, IF/IHC

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all SMAD6 Primary Antibodies »

Product Specifications

Immunogen

Synthetic peptide made to an internal portion of human SMAD6 (between amino acids 30-150) [UniProt O43541].

Reactivity Notes

Cross reacts with New World Monkey. Predicted to react with Equine. Mouse reactivity reported from a customer review.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for SMAD6 Antibody - BSA Free

Western Blot: SMAD6 Antibody [NB100-56440]

Western Blot: SMAD6 Antibody [NB100-56440]

Western Blot: SMAD6 Antibody [NB100-56440] - Analysis in mouse adult lung tissue. WB image submitted by a verified customer review.
Immunocytochemistry/ Immunofluorescence: SMAD6 Antibody [NB100-56440]

Immunocytochemistry/ Immunofluorescence: SMAD6 Antibody [NB100-56440]

Immunocytochemistry/Immunofluorescence: SMAD6 Antibody [NB100-56440] - HepG2 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton X-100. The cells were incubated with anti-SMAD6 (NB100-56440) at a 1:100 dilution overnight at 4C and detected with an anti-rabbit DyLight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse DyLight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.
Immunohistochemistry-Paraffin: SMAD6 Antibody [NB100-56440]

Immunohistochemistry-Paraffin: SMAD6 Antibody [NB100-56440]

Immunohistochemistry-Paraffin: SMAD6 Antibody [NB100-56440] - SMAD6 was detected in immersion fixed paraffin-embedded sections of human lung using Rabbit Anti-Human SMAD6 polyclonal Antibody (Catalog # NB100-56440) at 5 ug/ml for 1 hour at room temperature followed by incubation with the Anti-Rabbit IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC003). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Nuclear staining was localized to the pneumocytes.
Western Blot: SMAD6 Antibody [NB100-56440]

Western Blot: SMAD6 Antibody [NB100-56440]

Western Blot: SMAD6 Antibody [NB100-56440] - Analysis using SMAD6 antibody. Lysate from Jurkat cells probed with SMAD6 antibody at 2 ug/ml.
Simple Western: SMAD6 Antibody [NB100-56440]

Simple Western: SMAD6 Antibody [NB100-56440]

Simple Western: SMAD6 Antibody [NB100-56440] - Simple Western lane view shows a specific band for SMAD6 in 0.5 mg/ml of HepG2 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Applications for SMAD6 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1 - 5 ug/mL. Use reported in scientific literature (PMID 20032053)

Immunohistochemistry

5 - 10 ug/mL

Immunohistochemistry-Paraffin

5 - 10 ug/mL. Use reported in scientific literature (PMID 20032053)

Simple Western

1:50

Western Blot

0.5 - 2 ug/mL. Use reported by customer review
Application Notes
A 52 kDa band is observed. Use of this antibody in IHC-P and ICC-IF applications is cited by Hogg et al 2010 in Endocrinology 151:1247-56 (PMID: 20032053) and this publication has documented protocols for these applications.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in HepG2 lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:50, apparent MW was 54 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 2 reviews rated 5 using NB100-56440 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: SMAD6

SMADs are members of the MAD-related family of molecules. MAD-related proteins are a family of intracellular proteins that are essential components in the signaling pathways of the serine/threonine kinase receptors of the transforming growth factor beta superfamily. SMADs can be divided into receptor-regulated SMADs (R-SMADs: SMAD1, SMAD2, SMAD5, SMAD8 and SMAD9), common-mediator SMAD (co-SMAD: SMAD4), and inhibitory SMADs (I-SMADs: SMAD6 and SMAD7). SMAD1, SMAD5, SMAD8 and SMAD9 have high degrees of homology and antibodies are available that recognize sequences common to all of them. SMAD8 and SMAD9 are typically used as alternate names for one another in the literature. Human SMAD1 is a 465 amino acid protein; GenBank Accession No. AAP36050.1. Human SMAD2 is a 467 amino acid protein; GenBank Accession No. AAC51918.1 Human SMAD3 is a 425 amino acid protein; GenBank Accession No. NP_005893.1 Human SMAD4 is a 552 amino acid protein GenBank Accession No. NP_005350.1. Human SMAD5 is a 465 amino acid protein; GenBank Accession No. AAC50791.1. Human SMAD6 is a 496 amino acid protein; GenBank Accession No. AAH12986.1 Human SMAD7 is a 426 amino acid protein; GenBank Accession No. AAB81354.1 Mouse SMAD8 is a 430 amino acid protein; GenBank Accession No. AAN85445.1 Human SMAD9 is a 430 amino acid protein; GenBank Accession No. NP_005896.1

Alternate Names

hSMAD6, HsT17432, MAD homolog 6, MADH6SMAD, mothers against DPP homolog 6 (Drosophila), MADH7, mothers against decapentaplegic homolog 6, Mothers against decapentaplegic, drosophila, homolog of, 6, Mothers against DPP homolog 6, SMAD 6, SMAD family member 6MAD, mothers against decapentaplegic homolog 6 (Drosophila), SMAD, mothers against DPP homolog 6, Smad6

Entrez Gene IDs

17130 (Mouse); 367100 (Rat)

Gene Symbol

SMAD6

UniProt

O43541 (Human)

Additional SMAD6 Products

Product Documents for SMAD6 Antibody - BSA Free

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

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Product Specific Notices for SMAD6 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for SMAD6 Antibody - BSA Free

Customer Reviews for SMAD6 Antibody - BSA Free (2)

5 out of 5
2 Customer Ratings
5 Stars
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1 Stars
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Customer Images

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  • SMAD6 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Lung tissue
    Species: Mouse
    Verified Customer | Posted 08/02/2021
    Analysis in mouse lung tissue.
    SMAD6 Antibody - BSA Free NB100-56440
  • SMAD6 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Adult Mouse Lung Tissue
    Species: Mouse
    Verified Customer | Posted 06/10/2019
    SMAD6 Antibody - BSA Free NB100-56440

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Protocols

View specific protocols for SMAD6 Antibody - BSA Free (NB100-56440):

Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes (keep slides in the sodium citrate buffer at all times).

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in PBS for 5 minutes.
3. Block each section with 100-400 ul blocking solution (1% BSA in PBS) for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4 C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul HRP polymer conjugated secondary antibody. Incubate 30 minutes at room temperature.
7. Wash sections three times in wash buffer for 5 minutes each.
8. Add 100-400 ul DAB substrate to each section and monitor staining closely.
9. As soon as the sections develop, immerse slides in deionized water.
10. Counterstain sections in hematoxylin.
11. Wash sections in deionized water two times for 5 minutes each.
12. Dehydrate sections.
13. Mount coverslips.

Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10-25 ug of total protein per lane.
2. Transfer proteins to PVDF membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain the membrane with Ponceau S (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot TBS -0.05% Tween 20 (TBST).
5. Block the membrane in 5% Non-fat milk in TBST (blocking buffer) for at least 1 hour.
6. Wash the membrane in TBST three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate overnight at 4C with gentle rocking.
8. Wash the membrane in TBST three times for 10 minutes each.
9. Incubate the membrane in diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) for 1 hour at room temperature.
10. Wash the blot in TBST three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturer's instructions.

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