Antibodies, Proteins, Kits and Reagents for Life Science

298 185 ₸ Product size

100 µl 298 185 ₸

Order now and get it on Tuesday March 09, 2021

Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR15010] to Sortilin/NT3
Suitable for: IHC-P, WB
Knockout validated
Reacts with: Mouse, Human

Product name

Anti-Sortilin/NT3 antibody [EPR15010]
See all Sortilin/NT3 primary antibodies
Description

Rabbit monoclonal [EPR15010] to Sortilin/NT3
Host species

Rabbit

Tested Applications & Species

Application	Species
IHC-P	Rat Human
WB	Mouse Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: SW480 whole cell lysate (ab3957), SW40, HeLa and SH-SY5Y cell lysates. IHC-P: Human brain 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.

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

EPR15010
Isotype

IgG
Research areas

Western blot - Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

All lanes : Anti-Sortilin/NT3 antibody [EPR15010] (ab188586) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : SORT1 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 92 kDa
Observed band size: 100 kDa
why is the actual band size different from the predicted?

Lanes 1- 2: Merged signal (red and green). Green - ab188586 observed at 100 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab188586 was shown to react with Sortilin/NT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab264772 (knockout cell lysate ab257696) was used. Wild-type HeLa and SORT1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab188586 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labeling Sortilin/NT3 with ab188586 at 1/250 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Sortilin/NT3 knockout HAP1 cell lysate (20 µg)
Lane 3: SW480 cell lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)

Lanes 1 - 4: Merged signal (red and green). Green - ab188586 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab188586 was shown to react with Sortilin/NT3 in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when Sortilin/NT3 knockout samples were examined. Wild-type and Sortilin/NT3 knockout samples were subjected to SDS-PAGE. ab188586 and ab8245 (loading control to GAPDH) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

All lanes : Anti-Sortilin/NT3 antibody [EPR15010] (ab188586) at 1/1000 dilution

Lane 1 : SH-SY5Y whole cell lysate
Lane 2 : Human hippocampus tissue lysate
Lane 3 : Human fetal brain tissue lysate
Lane 4 : Mouse brain tissue lysate
Lane 5 : 3T3-L1 whole cell lysate
Lane 6 : Rat brain tissue lysate

Lysates/proteins at 15 µg per lane.

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

Predicted band size: 92 kDa
Observed band size: 92 kDa

Exposure time: 30 seconds

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

All lanes : Anti-Sortilin/NT3 antibody [EPR15010] (ab188586) at 1/2000 dilution

Lane 1 : SW480 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : SH-SY5Y cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 92 kDa
Anti-Sortilin/NT3 antibody [EPR15010] (ab188586)

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