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Phospho-Smad5-S465 Rabbit mAb (AP1411)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Mouse, Rat

Western blot analysis of lysates from NIH/3T3 cells, using Phospho-Smad5-S465 Rabbit mAb (AP1411) at 1:2000 dilution. NIH/3T3 cells were treated by BMP4(50ng/ml) at 37℃ for 6 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 180s.

Western blot analysis of lysates from C6 cells, using Phospho-Smad5-S465 Rabbit mAb (AP1411) at 1:2000 dilution. C6 cells were treated by MG132(50 μM) and TGFβ at 37℃ for 90 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 180s.

Overview

Product name	Phospho-Smad5-S465 Rabbit mAb
Catalog No.	AP1411
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC59204

Background

The protein encoded by this gene is involved in the transforming growth factor beta signaling pathway that results in an inhibition of the proliferation of hematopoietic progenitor cells. The encoded protein is activated by bone morphogenetic proteins type 1 receptor kinase, and may be involved in cancer. Alternative splicing results in multiple transcript variants.

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S465 of human Smad5.
Sequence	Email for sequence
Gene ID
Swiss Prot
Synonyms	DWFC; JV5-1; MADH5; Phospho-Smad5-S465
Calculated MW	52kDa
Observed MW	58kDa

Applications

Reactivity	Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:1000 - 1:5000
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting
Positive samples	NIH/3T3 treated by BMP4, C6 treated by TGFβ and MG132
Cellular location	Cytoplasm, Nucleus