Name: Anti-SATB2 antibody [EPNCIR130A]
Brand: RabMAb
SKU: ab92446
Rating: 4 (7 reviews)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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 (Intra)
Knockout validated
Reacts with: Mouse, Rat, Human

Alexa Fluor® 488 Alexa Fluor® 594 Alexa Fluor® 647 APC Carrier Free PE

Product name

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

Rabbit monoclonal [EPNCIR130A] to SATB2
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
Unsuitable for: IP
Species reactivity

Reacts with: Mouse, Rat, Human
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 (intra): 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
- Neuroscience
- Development

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab92446 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB	(1)	1/1000 - 1/5000. Predicted molecular weight: 81 kDa.
IHC-P		1/150 - 1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
ICC/IF	(1)	1/100.
Flow Cyt (Intra)		1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Notes
WB 1/1000 - 1/5000. Predicted molecular weight: 81 kDa.
IHC-P 1/150 - 1/300. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols.
ICC/IF 1/100.
Flow Cyt (Intra) 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Application notes

Is unsuitable for IP.

Function

Binds to DNA, at nuclear matrix- or scaffold-associated regions. Thought to recognize the sugar-phosphate structure of double-stranded DNA. Transcription factor controlling nuclear gene expression, by binding to matrix attachment regions (MARs) of DNA and inducing a local chromatin-loop remodeling. Acts as a docking site for several chromatin remodeling enzymes and also by recruiting corepressors (HDACs) or coactivators (HATs) directly to promoters and enhancers. Required for the initiation of the upper-layer neurons (UL1) specific genetic program and for the inactivation of deep-layer neurons (DL) and UL2 specific genes, probably by modulating BCL11B expression. Repressor of Ctip2 and regulatory determinant of corticocortical connections in the developing cerebral cortex. May play an important role in palate formation. Acts as a molecular node in a transcriptional network regulating skeletal development and osteoblast differentiation.
Tissue specificity

High expression in adult brain, moderate expression in fetal brain, and weak expression in adult liver, kidney, and spinal cord and in select brain regions, including amygdala, corpus callosum, caudate nucleus, and hippocampus.
Involvement in disease

Note=Chromosomal aberrations involving SATB2 are found in isolated cleft palate. Translocation t(2;7); translocation t(2;11).
Defects in SATB2 are a cause of cleft palate isolated (CPI) [MIM:119540]. A congenital fissure of the soft and/or hard palate, due to faulty fusion. Isolated cleft palate is not associated with cleft lips. Some patients may manifest other craniofacial dysmorphic features, mental retardation, and osteoporosis.
Note=A chromosomal aberration involving SATB2 is found in a patient with classical features of Toriello-Carey syndrome. Translocation t(2;14)(q33;q22).
Sequence similarities

Belongs to the CUT homeobox family.
Contains 2 CUT DNA-binding domains.
Contains 1 homeobox DNA-binding domain.
Post-translational
modifications

Sumoylated by PIAS1. Sumoylation promotes nuclear localization, but represses transcription factor activity.
Cellular localization

Nucleus matrix.
Target information above from: UniProt accession Q9UPW6 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 23314 Human
- Entrez Gene: 212712 Mouse
- Entrez Gene: 501145 Rat
- Omim: 608148 Human
- SwissProt: Q9UPW6 Human
- SwissProt: Q8VI24 Mouse
- Unigene: 145599 Mouse
Alternative names
- DNA binding protein SATB2 antibody
- DNA-binding protein SATB2 antibody
- FLJ21474 antibody
- FLJ32076 antibody
- GLSS antibody
- KIAA1034 antibody
- MGC119474 antibody
- MGC119477 antibody
- SATB family member 2 antibody
- SATB homeobox 2 antibody
- SATB2 antibody
- SATB2_HUMAN antibody
- Special AT rich sequence binding protein 2 antibody
- Special AT-rich sequence-binding protein 2 antibody
see all

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 (Intracellular) - Anti-SATB2 antibody [EPNCIR130A] (ab92446)

Intracellular 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)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab92446? Please let us know so that we can cite the reference in this datasheet.

ab92446 has been referenced in 43 publications.

Feurle P et al. SATB2-LEMD2 interaction links nuclear shape plasticity to regulation of cognition-related genes. EMBO J 40:e103701 (2021). PubMed: 33319920
Wang C et al. ApoE-Isoform-Dependent SARS-CoV-2 Neurotropism and Cellular Response. Cell Stem Cell 28:331-342.e5 (2021). PubMed: 33450186
Thornton CA et al. Spatially mapped single-cell chromatin accessibility. Nat Commun 12:1274 (2021). PubMed: 33627658
Hu HB et al. LPA signaling acts as a cell-extrinsic mechanism to initiate cilia disassembly and promote neurogenesis. Nat Commun 12:662 (2021). PubMed: 33510165
Chang SH et al. MicroRNAs mediate precise control of spinal interneuron populations to exert delicate sensory-to-motor outputs. Elife 10:N/A (2021). PubMed: 33787491

View all Publications for this product

Customer reviews and Q&As

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1-10 of 13 Abreviews or Q&A

Application

Immunohistochemistry (Frozen sections)

Sample

Mouse Tissue sections (E14.5 whole embryo)

Specification

E14.5 whole embryo

Blocking step

Gelatin as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 0.25%

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Alain Chedotal

Verified customer

Submitted Nov 23 2020

Application

Immunocytochemistry

Sample

Human Cultured Cells (neurons derived from iPSCs)

Specification

neurons derived from iPSCs

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 20 2017

Application

Immunocytochemistry

Sample

Mouse Cultured Cells (cortical neurons)

Specification

cortical neurons

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 22°C

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 12 2016

Application

Immunohistochemistry (PFA perfusion fixed frozen sections)

Blocking step

BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 25°C

Antigen retrieval step

None

Sample

Mouse Tissue sections (E18 mouse brain)

Specification

E18 mouse brain

Permeabilization

Yes - 0.3% Triton X-100

Fixative

Paraformaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 11 2013

Application

Western blot

Sample

Human Cell lysate - whole cell (MDA-MB231, H1299, SHSY5Y and HEK293T cells)

Loading amount

30 µg

Specification

MDA-MB231, H1299, SHSY5Y and HEK293T cells

Gel Running Conditions

Reduced Denaturing (10 % PAGE gel)

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 02 2012

Application

Immunocytochemistry/ Immunofluorescence

Sample

Human Cell (MDA-MB231 cells)

Specification

MDA-MB231 cells

Fixative

Paraformaldehyde

Permeabilization

Yes - 0.1 % TritonX-100 for 5 minutes

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 02 2012

Question

Immunogen of ab92446?

Abcam community

Verified customer

Asked on Feb 21 2012

Answer

Thank you for your call yesterday.
I now heard back form the source of this antibody and I am very happy to confirm that we can provide a range of amino acids where the immunogen in a part of.
Unfortunately the exact sequence is proprietary and was not communicated to me.
The immunogen peptide maps between amino acids 240 and 275 of human SATB2 (UniProt: Q9UPW6).
I hope this information will be helpful to the interpretation of your data.

Abcam Scientific Support

Answered on Feb 21 2012

Question

Hi,

I am testing your antibodies ab51502 and ab92446 for WB and IP and would like to know the concentration of the antibody.

Also, is there a recommended IP protocol or preference of the 2 antibody for IP or WB?

Thank you!

Abcam community

Verified customer

Asked on Feb 16 2012

Answer

Thank you for contacting us. The concentration of ab51502 is 0.1 mg/ml and should be used at a dilution of 1/100 for both IP and WB. Regarding ab92445, this is a tissue culture supernatant antibody so the concentration is not determined. Generally concentrations are only determined for purified antibodies as they are not relevant for other types that may contains proteins other than the protein of interest. This product should be used at a dilution of 1/10-1/100 for IP and 1/1000-1/10000 for WB. We do not test our antibodies side by side and cannot say which works "better" for a particular application but we do guarantee them both to work in IP and WB.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Feb 16 2012

Question

As a customer, I have just recently purchased several antibodies from Abcam, most of them are working very nicely. But there is one antibody, Rabbit monoclonal [EPNCIR130A] to SATB2 (Cat. ab92446, Lot. GR7835-5) does NOT perform as shown on the datasheet. We did our immunocytochemistry right after the SATB2 antibody arrived, there is no any nuclear staining (where the protein should localize), we could only visualize a lot of aggregates with fluorescence distributing everywhere not in cells. It seems very obvious the SATB2 antibody had been forming aggregates before and/or during the antibody had been delivered. Since this is a very important piece of data for our study, I am now writing to ask for Replacement of this antibody ab92446.

Abcam community

Verified customer

Asked on Nov 08 2011

Answer

Thank you for contacting us. I am very sorry to hear that you are having problems with this antibody. Could you please let me know a few more experimental details (for our records): 1) What sample type was used and what is the species of the samples? 2) What antibody dilution was tried? 3) Which blocking reagent was used? 4) Which secondary antibody/detection system was employed? Also, please let me know your order number. If the item was purchased within the last 6 months, I'd be happy to discuss a replacement with you. Please let me know the requested information. I look forward to hear back from you and assist in resolving this case for you.

Abcam Scientific Support

Answered on Nov 08 2011

Question

Thanks for your help. I did finally get a signal after letting the exposure go for much longer, so it looks like it worked. I would like to get a 2nd SATB2 antibody to use to confirm the IP I plan to do. Can you recommend another SATB2 antibody that has been successful in Western analysis or IP?

Abcam community

Verified customer

Asked on Nov 08 2011

Answer

Thank you for your email. I am very glad to hear that you obtained the desired results. As for another SATB2 antibody that is suitable for IP/WB, we have also ab92446: https://www.abcam.com/SATB2-antibody-EPNCIR130A-ab92446.html It was tested in IP and WB and several other applications, as well as in human, mouse and rat samples. The antibody you are currently using is also suitable for IP: ab51502 Other antibodies tested in WB and human samples are: ab69995, ab69998 and ab119118. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Nov 08 2011

1-10 of 13 Abreviews or Q&A

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (43)

Customer reviews and Q&As

Immunohistochemistry (Frozen sections) abreview for Anti-SATB2 antibody [EPNCIR130A]

Immunocytochemistry abreview for Anti-SATB2 antibody [EPNCIR130A]

Immunocytochemistry abreview for Anti-SATB2 antibody [EPNCIR130A]

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-SATB2 antibody [EPNCIR130A]

Western blot abreview for Anti-SATB2 antibody [EPNCIR130A]

Immunocytochemistry/ Immunofluorescence abreview for Anti-SATB2 antibody [EPNCIR130A]

Post-translational
modifications