Antibodies, Proteins, Kits and Reagents for Life Science

291 484 ₸ Product size

100 µl 291 484 ₸

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Anti-SMYD3 antibody [EPR19311] (ab199361)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR19311] to SMYD3
Suitable for: WB, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-SMYD3 antibody [EPR19311]
See all SMYD3 primary antibodies
Description

Rabbit monoclonal [EPR19311] to SMYD3
Host species

Rabbit
Tested Applications & Species

Application Species
IP
Human

WB
Mouse

Rat

Human

See all applications and species data
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal kidney and fetal brain lysates; wild-type HAP1, HeLa, MCF7, HEK-293, T-47D, HCT 116, LLC, C6, RAW 264.7, PC-12 and NIH/3T3 whole cell lysates; Mouse brain, kidney and spleen lysates; Rat brain and kidney lysates. IP: 293T whole cell lysate.
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.

Application	Species
IP	Human
WB	Mouse Rat Human

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
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

EPR19311
Isotype

IgG
Research areas

Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)

All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SMYD3 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : MCF7 whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 49 kDa
Observed band size: 49 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab199361 observed at 49 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab199361 was shown to react with SMYD3 in HAP1 wild-type cells in Western blot. Loss of signal was observed when SMYD3 knockout sample was used. HAP1 wild-type and SMYD3 knockout whole cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with ab199361 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)

All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 49 kDa
Observed band size: 45 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)

All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution

Lane 1 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : T-47D (Human ductal breast epithelial tumor cell line) whole cell lysate
Lane 4 : HCT 116 (Human colorectal carcinoma cell line) whole cell lysate
Lane 5 : LLC (Mouse lung carcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 49 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-SMYD3 antibody [EPR19311] (ab199361)

All lanes : Anti-SMYD3 antibody [EPR19311] (ab199361) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Mouse spleen lysate
Lane 4 : Rat brain lysate
Lane 5 : Rat kidney lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
Lane 8 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
Lane 9 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 49 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)

SMYD3 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199361 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab199361 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: 293T whole cell lysate, 10µg (Input).

Lane 2: ab199361 IP in 293T whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A ]-Isotype Control (ab172730) instead of ab199361 in 293T whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.
Immunoprecipitation - Anti-SMYD3 antibody [EPR19311] (ab199361)

SMYD3 was immunoprecipitated from 1mg of 293T (Human epithelial cell line from embryonic kidney) whole cell lysate with ab199361 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab199361 at 1/1000 dilution (Panel A) or ab183498 at 1/1000 dilution (Panel B).

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1: 293T whole cell lysate, 10µg (Input).

Lane 2: ab199361 IP in 293T whole cell lysate.

Lane 3: Rabbit IgG,monoclonal [EPR25A]-Isotype Control (ab172730) instead of ab199361 in 293T whole cell lysate.

Blocking and dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: 3 minutes.

The product was validated with ab183498, which is specific to SMYD3.
Anti-SMYD3 antibody [EPR19311] (ab199361)

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