Antibodies, Proteins, Kits and Reagents for Life Science

361 843 ₸ Product size

100 µl 361 843 ₸ 10 µl 144 067 ₸

Order now and get it on Tuesday March 02, 2021

Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPNCIR130A] to SATB2
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-SATB2 antibody [EPNCIR130A]
See all SATB2 primary antibodies
Description

Rabbit monoclonal [EPNCIR130A] to SATB2
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Mouse Human

See all applications and species data

Immunogen

Synthetic peptide within Human SATB2. The exact sequence is proprietary.
Database link: Q9UPW6
Positive control
- WB: HT-1080, SW1353, MCF7 and Saos-2 cell lysates; Rat and mouse brain and human fetal brain tissue lysates; Wild-type HAP1 whole cell lysate. IHC-P: Human cerebral cortex tissue; Mouse brain tissue. ICC/IF: SH-SY5Y cells. Flow Cyt: SH-SY5Y cells. IHC-Fr: Mouse brain tissue.
General notes
This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of John Niederhuber. View antibodies from NCI Center for Cancer Research Collaboration.
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: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPNCIR130A
Isotype

IgG
Research areas

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling SATB2 with purified ab92446 at 1/150. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Western blot - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

All lanes : Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SATB2 knockout HAP1 whole cell lysate

Lysates/proteins at 20 µg per lane.

Predicted band size: 81 kDa

Lanes 1 - 2: Merged signal (red and green). Green - ab92446 observed at 83 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab92446 was shown to specifically react with SATB2 in wild-type HAP1 cells as signal was lost in SATB2 knockout cells. Wild-type and SATB2 knockout samples were subjected to SDS-PAGE. Ab92446 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.
Immunocytochemistry/ Immunofluorescence - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y (human neuroblastoma cell line from bone marrow) cells labelling SATB2 (green) with purified ab92446 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.

Control: primary antibody (1/100) and secondary antibody ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Flow Cytometry - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Flow Cytometry analysis of SH-SY5Y cells (human neuroblastoma cell line from bone marrow) labeling SATB2 with purified ab92446 at 1/150 dilution(10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488)(ab150077)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black)(ab172730) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.
Western blot - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution (purified) + SW1353 (human chondrosarcoma cell line) cell lysate at 20 µg

Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 81 kDa
Observed band size: 83 kDa
why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

All lanes : Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution (purified)

Lane 1 : Saos-2 (human osteosarcoma cell line) cell lysate
Lane 2 : Human fetal brain tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 81 kDa
Observed band size: 83 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

All lanes : Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution (purified)

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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 81 kDa
Observed band size: 83 kDa why is the actual band size different from the predicted?

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

All lanes : Anti-SATB2 antibody [EPNCIR130A] (ab92446) at 1/1000 dilution (unpurified)

Lane 1 : HT1080 (human fibrosarcoma cell line) cell lysate
Lane 2 : SW1353 (human chondrosarcoma cell line) cell lysate
Lane 3 : MCF7 (human breast adenocarcinoma cell line) cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 81 kDa
Anti-SATB2 antibody [EPNCIR130A] (ab92446)

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