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WB analysis using 87035-1-RR (same clone as 87035-1-PBS)
Various lysates were subjected to SDS PAGE followed by western blot with 87035-1-RR (SMAD3 antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 87035-1-PBS in a different storage buffer formulation.
Sandwich ELISA sample test of MP02853-2
The mean SMAD3 concentration was determined to be 503.2 ng/mL in A431 cell extract based on a 1.2 mg/mL extract load, 253.4 ng/mL in A549 cell extract based on a 1.2 mg/mL extract load, 709.6 ng/mL in SH-SY5Y cell extract based on a 1.2 mg/mL extract load, 646.7 ng/mL in Raji cell extract based on a 1.2 mg/mL extract load and 750.4 ng/mL in NIH/3T3 cell extract based on a 1.2 mg/mL extract load.
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The mean SMAD3 concentration was determined to be 503.2 ng/mL in A431 cell extract based on a 1.2 mg/mL extract load, 253.4 ng/mL in A549 cell extract based on a 1.2 mg/mL extract load, 709.6 ng/mL in SH-SY5Y cell extract based on a 1.2 mg/mL extract load, 646.7 ng/mL in Raji cell extract based on a 1.2 mg/mL extract load and 750.4 ng/mL in NIH/3T3 cell extract based on a 1.2 mg/mL extract load.
Sandwich ELISA sample test of MP02853-2
Serum of sixteen individual healthy human donors was measured. The SMAD3 concentration of detected samples was determined to be 128.2 pg/mL with a range of 31.3-281.8 pg/mL.
Sandwich ELISA standard curve of MP02853-2
Sandwich ELISA standard curve of MP02853-2, Human SMAD3 Recombinant Matched Antibody Pair - PBS only. 87035-4-PBS was coated to a plate as the capture antibody and incubated with serial dilutions of standard Ag32623. 87035-1-PBS was HRP conjugated as the detection antibody. Range: 46.9-3000 pg/mL.
IHC staining of mouse prostate using 87035-1-RR (same clone as 87035-1-PBS)
Immunohistochemical analysis of paraffin-embedded mouse prostate tissue slide using 87035-1-RR (SMAD3 antibody) at dilution of 1:200 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 87035-1-PBS in a different storage buffer formulation.
IF Staining of HEK-293 using 87035-1-RR (same clone as 87035-1-PBS)
Immunofluorescent analysis of (4% PFA) fixed HEK-293 cells using SMAD3 antibody (87035-1-RR, Clone: 251981H8 ) at dilution of 1:1000 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2), CL594-Phalloidin (red). This data was developed using the same antibody clone with 87035-1-PBS in a different storage buffer formulation.
Affinity and Kinetic Characterization of 87035-1-RR
Biolayer interferometry (BLl) kinetic assays of 87035-1-RR against Human SMAD3 were performed. The affinity constant is 1.26 nM.

SMAD3 Recombinant monoclonal antibody, PBS Only (Detector)

Name: SMAD3 Recombinant monoclonal antibody, PBS Only (Detector)
Brand: Proteintech
SKU: 87035-1-PBS

SMAD3 Uni-rAb^® Recombinant Antibody for WB, IF/ICC, Sandwich ELISA, Indirect ELISA

Cat No. 87035-1-PBS

Clone No.251981H8

Review Product

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IF/ICC, Sandwich ELISA, Indirect ELISA

Synonyms

hMAD 3, hMAD-3, hSMAD3, HSPC193, HsT17436

Formulation: PBS Only

PBS, Azide, Glycerol

PBS Only

Conjugate: Unconjugated

Size/Concentration:

-/ -

Freight/Packing: -

Quantity

- +

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Overview

Tested Applications
Product Information

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MSDS SDS

Product Information

87035-1-PBS targets SMAD3 as part of a matched antibody pair:

MP02853-2: 87035-4-PBS capture and 87035-1-PBS detection (validated in Sandwich ELISA)

Unconjugated rabbit recombinant monoclonal antibody in PBS only (BSA and azide free) storage buffer at a concentration of 1 mg/mL, ready for conjugation. Created using Proteintech’s proprietary in-house recombinant technology. Recombinant production enables unrivalled batch-to-batch consistency, easy scale-up, and future security of supply.

This conjugation ready format makes antibodies ideal for use in many applications including: ELISAs, multiplex assays requiring matched pairs, mass cytometry, and multiplex imaging applications.Antibody use should be optimized by the end user for each application and assay.

Tested Reactivity	human, mouse, rat
Host / Isotype	Rabbit / IgG
Class	Recombinant
Type	Antibody
Immunogen	CatNo: Ag32623 Product name: Recombinant human SMAD3 protein Source: e coli.-derived, PGEX-4T Tag: GST Domain: 145-260 aa of NM_005902 Sequence: EIPAEFPPLDDYSHSIPENTNFPAGIEPQSNIPETPPPGYLSEDGETSDHQMNHSMDAGSPNLSPNPMSPAHNNLDLQPVTYCEPAFWCSISYYELNQRVGETFHASQPSMTVDGF Predict reactive species
Full Name	SMAD family member 3
Calculated Molecular Weight	48 kDa
Observed Molecular Weight	52 kDa
GenBank Accession Number	NM_005902
Gene Symbol	SMAD3
Gene ID (NCBI)	4088
Conjugate	Unconjugated
Form	Liquid
Purification Method	Protein A purification
UNIPROT ID	P84022
Storage Buffer	PBS only, pH 7.3.
Storage Conditions	Store at -80°C.

Background Information

SMAD3, also named as hMAD 3 or Mad3, is a 425 amino acid protein, which contains one MH1 domain and one MH2 domain. SMAD3 localizes in the nucleus and cytoplasm. SMAD3 plays an essential role in development and maintenance of self-tolerance and is a critical mediator of the TGFB signaling pathway. SMAD3 is involved in TGFB dependent regulation of steroidogenesis and in T-cell response to TGFB.