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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [2]
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- Validations
- Western blot [3]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Flow cytometry [1]
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- Product number
- MA5-15682 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SMAD4 Monoclonal Antibody (4G1C6)
- Antibody type
- Monoclonal
- Antigen
- Purifed from natural sources
- Description
- MA5-15682 targets SMAD4 in FACS, IF, IHC, and WB applications and shows reactivity with Human samples. The MA5-15682 immunogen is purified recombinant fragment of human SMAD4 expressed in E. Coli.
- Antibody clone number
- 4G1C6
- Concentration
- Conc. Not Determined
Submitted references Operation of the Atypical Canonical Bone Morphogenetic Protein Signaling Pathway During Early Human Odontogenesis.
MiR-34a-3p alters proliferation and apoptosis of meningioma cells in vitro and is directly targeting SMAD4, FRAT1 and BCL2.
Hu X, Lin C, Ruan N, Huang Z, Zhang Y, Hu X
Frontiers in physiology 2022;13:823275
Frontiers in physiology 2022;13:823275
MiR-34a-3p alters proliferation and apoptosis of meningioma cells in vitro and is directly targeting SMAD4, FRAT1 and BCL2.
Werner TV, Hart M, Nickels R, Kim YJ, Menger MD, Bohle RM, Keller A, Ludwig N, Meese E
Aging 2017 Mar 23;9(3):932-954
Aging 2017 Mar 23;9(3):932-954
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of SMAD4 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPRMA515682_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of SMAD4 was performed by loading 30 µg of HeLa Wild Type (Lane 1), HeLa CAS9 (Lane 2) and HeLa SMAD4 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-SMAD4 Monoclonal Antibody (4G1C6) (Product # MA515682, 1:2000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to SMAD4. Uncharacterized band at ~ 46 kDa was observed in all the samples.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SMAD4 Monoclonal Antibody (4G1C6) (Product # MA5-15682) and a 60 kDa band corresponding to Mothers against decapentaplegic homolog 4, along with an uncharacterized band(*), was observed across positive cell lines (HCT 116 and A-431) and not across negative cell lines (HT-29 and SW480) tested. Whole cell extracts (30 µg lysate) of HCT 116 (Lane 1), A-431 (Lane 2), HT-29 (Lane 3) and SW480 (Lane 4) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SMAD4 using SMAD4 monoclonal antibody (Product # MA5-15682) in A431 (1), SK-N-SH (2), K562 (3), HepG2 (4) and HUVE12 (5) cell lysate.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of NIH/3T3 cells using SMAD4 monoclonal antibody (Product # MA5-15682) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of NIH/3T3 cells using SMAD4 monoclonal antibody (Product # MA5-15682) (Green). Blue: DRAQ5 fluorescent DNA dye. Red: actin filaments have been labeled with phalloidin.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of paraffin-embedded lung cancer tissues using SMAD4 monoclonal antibody (Product # MA5-15682) followed with DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometric analysis of K562 cells using SMAD4 monoclonal antibody (Product # MA5-15682) (green) and negative control (purple).
