bs-3418R
[Primary Antibody]
Smad1/5 (Ser463 + Ser465) Antibody
DATASHEET
Host: Rabbit
Target Protein: Smad1/5 Ser463 + Ser465
Specificity: This phosphorylation site is homologous in the listed cross reactive species at the specified location.
Modification Site: Ser463 + Ser465
Clonality: Polyclonal
Isotype: IgG
Entrez Gene: 4086, 4090
Source: KLH conjugated synthetic phosphopeptide derived from human Smad1/5 around the phosphorylation site of Ser463/465
Purification: Purified by Protein A.
Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Background:
The protein encoded by this gene 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. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein mediates the signals of the bone morphogenetic proteins (BMPs), which are involved in a range of biological activities including cell growth, apoptosis, morphogenesis, development and immune responses. In response to BMP ligands, this protein can be phosphorylated and activated by the BMP receptor kinase. The phosphorylated form of this protein forms a complex with SMAD4, which is important for its function in the transcription regulation. This protein is a target for SMAD-specific E3 ubiquitin ligases, such as SMURF1 and SMURF2, and undergoes ubiquitination and proteasome-mediated degradation. Alternatively spliced transcript variants encoding the same protein have been observed. [provided by RefSeq].
Size: 100ul
Concentration: 1ug/ul
Applications:
WB(1:300-5000)
ELISA(1:500-1000)
IHC-P(1:200-400)
IHC-F(1:100-500)
IF(IHC-P)(1:50-200)
IF(IHC-F)(1:50-200)
IF(ICC)(1:50-200)
ICC(1:100-500)
Predicted Molecular Weight: 52
Cross Reactive Species:
Mouse
Rat
Predicted Cross Reactive Species: Human
For research use only. Not intended for diagnostic or therapeutic use.
PRODUCT SPECIFIC PUBLICATIONS
- Guo LP et al. Smad signaling coincides with epithelial-mesenchymal transition in a rat model of intrauterine adhesion. Am J Transl Res. 2019 Aug 15;11(8):4726-4737. eCollection 2019.Read more>>
VALIDATION IMAGES
Mouse intestine lysates probed with Rabbit Anti-Smad1/5 (Ser463+Ser465) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:300 overnight at 4˚C. Followed by a conjugated secondary antibody (bs-0295G-HRP) at 1:5000 for 90 min at RT.
Lane 1: Mouse Lymph node lysates; Lane 2: Mouse Uterus lysates probed with Smad1/5 (Ser463 + Ser465) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad1/5 (Ser463 + Ser465) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:400 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with Smad1/5 (Ser463 + Ser465) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat heart); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat skin); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Smad1/5 (Ser463 + Ser465)) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Lane 1: Mouse Cerebrum tissue lysates; Lane 2: Mouse Heart tissue lysates; Lane 3: Mouse Testis tissue lysates; Lane 4: Mouse Skin tissue lysates; Lane 5: Mouse Kidney tissue lysates; Lane 6: Rat Cerebrum tissue lysates; Lane 7: Rat Testis tissue lysates; Lane 8: Rat Skin tissue lysates; Lane 9: Rat Kidney tissue lysates; Lane 10: Human Huvec cell lysates; Lane 11: Human A549 cell lysates; Lane 12: Human HeLa cell lysates; Lane 13: Human HT1080 cell lysates; Lane 14: Human A431 cell lysates probed with Phospho-Smad1/5 (Ser463 + Ser465) Polyclonal Antibody, Unconjugated (bs-3418R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at room temperature for 60 min.