PA5-34806

antibody from Invitrogen Antibodies

Targeting: SMAD4 DPC4, MADH4

Provider product page for PA5-34806

Western blot

Immunocytochemistry

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [3]
Comments [0]
Validations
Immunocytochemistry [3]
Other assay [2]

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Product number: PA5-34806 - Provider product page
Provider: Invitrogen Antibodies
Product name: SMAD4 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant protein fragment
Description: Recommended positive controls: 293T, A431, HeLa, HepG2, Neuro 2A, C8D30, NIH-3T3, Raw264.7, C2C12, PC-12, Rat2, DDDDK-tagged SMAD4-transfected 293T. Predicted reactivity: Mouse (100%), Rat (100%), Pig (100%), Sheep (100%), Rhesus Monkey (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.58 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

SMAD4 protein structure - PA5-34806 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of SMAD4 was performed using 70 % confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with SMAD4 Polyclonal Antibody (PA5-34806) at 5 µg/mL in 0.1% BSA and incubated overnight at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear staining. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of SMAD4 was performed in SK-N-SH cells fixed in 4% paraformaldehyde at RT for 15 min. Green: SMAD4 Polyclonal Antibody (Product # PA5-34806) diluted at 1:500. Blue: Hoechst 33342 staining. Scale bar = 10 µm.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of SMAD4 was performed in HeLa cells fixed in 4% paraformaldehyde at RT for 15 min. Green: SMAD4 Polyclonal Antibody (Product # PA5-34806) diluted at 1:500. Blue: Hoechst 33342 staining.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 9 Smad4 regulates diencephalic astrocyte proliferation and neurosphere formation A Reactome pathway analysis for all 174 transcription factor genes with higher expression in DIE (upper) and all 205 genes higher in CTX GM (lower) astrocytes. B t-SNE visualization of DIE astrocytes isolated by ACSA-2 MACS (Fig 1A ) showing Smad4 expression (scaled log-normalized read counts). C Example of a Smad4/GFP double-positive astrocyte (indicated by arrowhead) in a single optical section of DIE from 3-mo-old GLAST CreERT2 /eGFP mice 21 dpt immunostained as indicated on top of the panel. Orthogonal projections of the area highlighted in left panel of C shown at higher magnification in the right panel. Scale bars: 15 um (left panel) and 10 um (right panel). D Example of a proliferating (PCNA + , arrowhead) astrocyte in a single optical section of DIE from 3-mo-old GLAST CreERT2 /eGFP mice 21 dpt immunostained as indicated on top with single channels shown in middle and right panels. Scale bars: 15 um. E Histogram depicting percentage of PCNA-positive cells of all GFP-positive cells in the DIE of the genotype indicated on the x-axis ( n = 6 animals per genotype; 5 ROI covering the diencephalon on a total of 3 slides were analysed per animal; PCNA-negative and PCNA-positive cells comparing wt/wt vs. fl/fl animals, adjusted P -value = 0.002). F, G Histograms depicting percentage of the neurosphere-generating cells obtained from the DIE (F) or SEZ (G) of GLAST CreERT2 /Smad4 fl/fl /eGFP

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1 Relative expression (median) of Smad4 gene and protein in addition to the protein localisation by IHC in the FFPE ( n = 90 patients) and fresh ( n = 40 patients) cohorts of paired malignant and normal colonic tissues according to clinical stages and tumour sidedness (Scale bar = 15 mum; a = p < 0.05 compared with right-sided normal tissue from early cancer; b = p < 0.05 compared with early stage right-sided colon cancer; c = p < 0.05 compared with left-sided normal tissue from early cancer; d = p < 0.05 compared with early stage left-sided colon cancer; e = p < 0.05 compared with right-sided normal tissue from advanced cancer; f = p < 0.05 compared with advanced stage right-sided colon cancer and g = p < 0.05 compared with left-sided normal tissue from advanced cancer).