Antibodies, Proteins, Kits and Reagents for Life Science

328 339 ₸ Product size

100 µl 328 339 ₸

Order now and get it on Tuesday March 02, 2021

Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [MMC-1-104-3] to SMAD5 (phospho S463 + S465)
Suitable for: Dot blot, WB, IHC-P
Reacts with: Mouse, Rat, Human

Product name

Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3]
See all SMAD5 primary antibodies
Description

Rabbit monoclonal [MMC-1-104-3] to SMAD5 (phospho S463 + S465)
Host species

Rabbit
Specificity

This antibody may cross-react with Smad1 Phospho (pS463/465) and Smad9 Phospho (pS465/467).

Tested Applications & Species

Application	Species
IHC-P	Mouse Rat Human
WB	Mouse Human

See all applications and species data

Immunogen

corresponding to Human SMAD5 (phospho S463 + S465).
Positive control
- WB: HeLa cell lysate treated with BMP-4. IHC-P: Human breast carcinoma, colonic carcinoma, tonsil, hepatocellular carcinoma, cervical carcinoma and glioma tissues.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

MMC-1-104-3
Isotype

IgG
Research areas

Western blot - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

All lanes : Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698) at 1/5000 dilution (purified)

Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 52 kDa
Observed band size: 58 kDa
why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM /TBST.
Western blot - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

All lanes : Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698) at 1/10000 dilution (purified)

Lane 1 : HeLa whole cell lysate - untreated
Lane 2 : HeLa whole cell lysate - treated with Bone Morphogenetic protein-4

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 52 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM /TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue labelling SMAD5 (phospho S463 + S465) with purified ab92698 at a dilution of 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue labelling SMAD5 (phospho S463 + S465) with purified ab92698 at a dilution of 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling SMAD5 (phospho S463 + S465) with purified ab92698 at a dilution of 1/800. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Dot Blot - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Dot blot analysis of SMAD5 (pS463 + pS465) peptide (Lane 1), SMAD5 (pS465) peptide (Lane 2), SMAD5 (pS463) peptide (Lane 3) and SMAD5 non-phospho peptide (Lane 4) labelling SMAD5 (pS465) with purified ab92698 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Western blot - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

All lanes : Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698) at 1/10000 dilution

Lane 1 : Untreated HeLa (human cervix adenocarcinoma) whole cell lysates 20µg
Lane 2 : HeLa (human cervix adenocarcinoma) treated with Bone Morphogenetic Protein-4 (BMP-4) whole cell lysates 20µg
Lane 3 : HeLa (human cervix adenocarcinoma) treated with Bone Morphogenetic Protein-4 (BMP-4) and phosphatase, whole cell lysates 20µg.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 52 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of (1) human breast carcinoma and (2) human colonic carcinoma tissues labelling SMAD5 (phospho S463 + P465) with unpurified ab92698 at a dilution of 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human tonsil tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human hepatocellular carcinoma tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervical carcinoma tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling SMAD5 (phospho S463 + S465) with unpurified ab92698.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-SMAD5 (phospho S463 + S465) antibody [MMC-1-104-3] (ab92698)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com