Recombinant Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Product name

Anti-GMEB1 antibody [EPR23056-62]
See all GMEB1 primary antibodies
Description

Rabbit monoclonal [EPR23056-62] to GMEB1
Host species

Rabbit
Tested applications

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

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK-293T transfected with mouse GMEB1, HeLa, A549, RAW264.7 and PC-12 lysates. IHC-P: Human colon, Human spleen, Human tonsil, Mouse spleen and Rat spleen tissues. ICC/IF: HeLa and Neuro-2a cells. Flow Cyt (intra): HeLa and HEK-293T cells. IP: HeLa cells.
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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR23056-62
Isotype

IgG
Research areas

Alternative Versions
- Anti-GMEB1 antibody [EPR23056-62] - BSA and Azide free (ab267346)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab240646 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		1/1000. Detects a band of approximately 82 kDa (predicted molecular weight: 63 kDa).
IP		1/30.
Flow Cyt (Intra)		1/500.
ICC/IF		1/50.
IHC-P		1/1000 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Use at 1/1000 dilution for human and mouse, 1/2000 dilution for rat.

Notes
WB 1/1000. Detects a band of approximately 82 kDa (predicted molecular weight: 63 kDa).
IP 1/30.
Flow Cyt (Intra) 1/500.
ICC/IF 1/50.
IHC-P 1/1000 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Use at 1/1000 dilution for human and mouse, 1/2000 dilution for rat.

Function

Trans-acting factor that binds to glucocorticoid modulatory elements (GME) present in the TAT (tyrosine aminotransferase) promoter and increases sensitivity to low concentrations of glucocorticoids. Binds also to the transferrin receptor promoter. Essential auxiliary factor for the replication of parvoviruses.
Sequence similarities

Contains 1 SAND domain.
Cellular localization

Nucleus. Cytoplasm. May be also cytoplasmic.
Target information above from: UniProt accession Q9Y692 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 10691 Human
- Entrez Gene: 56809 Mouse
- Entrez Gene: 500558 Rat
- Omim: 604409 Human
- SwissProt: Q9Y692 Human
- SwissProt: Q9JL60 Mouse
- SwissProt: Q9QUZ8 Rat
- Unigene: 632373 Human
- Unigene: 103489 Mouse
- Unigene: 219045 Rat
see all
Alternative names
- DNA binding protein p96PIF antibody
- DNA-binding protein p96PIF antibody
- Glucocorticoid modulatory element binding protein 1 antibody
- Glucocorticoid modulatory element-binding protein 1 antibody
- GMEB 1 antibody
- GMEB-1 antibody
- Gmeb1 antibody
- GMEB1_HUMAN antibody
- P96PIF antibody
- Parvovirus initiation factor p96 antibody
- PIF 96 antibody
- PIF p96 antibody
- PIF96 antibody
see all

Western blot - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

All lanes : Anti-GMEB1 antibody [EPR23056-62] (ab240646) at 1/1000 dilution

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
Lane 2 : A549 (human lung carcinoma epithelial cell), whole cell lysate
Lane 3 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 63 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure times.

Lane 1: 26 seconds; Lane 2: 125 seconds; Lane 3: 169 seconds; Lane 4: 3 minutes.

The weak lower bands below 75kDa could be degraded GMEB1 fragments (PMID:10386584).
Western blot - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

All lanes : Anti-GMEB1 antibody [EPR23056-62] (ab240646) at 1/1000 dilution

Lane 1 : HEK-293T (human embryonic kidney) transfected with an empty vector (vector control), containing a myc-His-tag®, whole cell lysate
Lane 2 : HEK-293T transfected with mouse GMEB1 (WT)(aa1-562) expression vector containing a myc-His-tag®, 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: 63 kDa
Observed band size: 82 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 seconds.

The weak lower bands below 75kDa could be degraded GMEB1 fragments (PMID:10386584).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling GMEB1 with ab240646 at 1/2000 dilution (0.288 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on rat spleen. The section was incubated with ab240646 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling GMEB1 with ab240646 at 1/1000 dilution (0.576 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on mouse spleen (PMID: 10692587) is observed. The section was incubated with ab240646 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling GMEB1 with ab240646 at 1/1000 dilution (0.576 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on human tonsil (PMID: 16497673) is observed. The section was incubated with ab240646 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling GMEB1 with ab240646 at 1/1000 dilution (0.576 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on human spleen (PMID: 16497673) is observed. The section was incubated with ab240646 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling GMEB1 with ab240646 at 1/1000 dilution (0.576 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer). Nuclear staining on human colon (PMID: 16497673) is observed. The section was incubated with ab240646 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP Polymer).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunoprecipitation - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

GMEB1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab240646 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab240646 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.

Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug

Lane 2: ab240646 IP in HeLa whole cell lysate

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab240646 in HeLa whole cell lysate

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

Exposure time: 3 min.
Flow Cytometry (Intracellular) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HEK-293T (human embryonic kidney) transfected with mouse GMEB1 (WT) (aa1-562) expression vector containing a myc-His-tag® cells labelling GMEB1 with ab240646 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) (Left). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

Cells were stained with rabbit IgG (Left) or ab240646 (Right). Then stained with anti-myc tag conjugated to PE.
Flow Cytometry (Intracellular) - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling GMEB1 with ab240646 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730, Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunocytochemistry/ Immunofluorescence - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling GMEB1 with ab240646 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing mainly nuclear and weakly cytoplasmic staining in Neuro-2a cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-GMEB1 antibody [EPR23056-62] (ab240646)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling GMEB1 with ab240646 at 1/50 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (Green). Confocal image showing mainly nuclear and weakly cytoplasmic staining in HeLa cell line. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution.
Anti-GMEB1 antibody [EPR23056-62] (ab240646)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

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Key features and details

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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

Isotype control

Applications

The Abpromise guarantee

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

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