Anti-NDRG1 antibody [EPR5593] KO tested (ab124689)

Rabbit Recombinant Monoclonal NDRG1 antibody. Suitable for IHC-P, IP, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 27 publications.

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Images

Western blot - Anti-NDRG1 antibody [EPR5593] (AB124689), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	Flow Cyt (Intra)	WB	ICC/IF
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Tested	Expected	Expected	Tested	Expected
Rat	Tested	Expected	Expected	Tested	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250 - 1/500	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Mouse	Dilution info 1/250 - 1/500	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/250 - 1/500	Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Rat	Dilution info 1/10000 - 1/50000	Notes -
Species Mouse	Dilution info 1/10000 - 1/50000	Notes -
Species Human	Dilution info 1/10000 - 1/50000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100 - 1/250	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

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Target data

See full target information NDRG1

Function

Stress-responsive protein involved in hormone responses, cell growth, and differentiation. Acts as a tumor suppressor in many cell types. Necessary but not sufficient for p53/TP53-mediated caspase activation and apoptosis. Has a role in cell trafficking, notably of the Schwann cell, and is necessary for the maintenance and development of the peripheral nerve myelin sheath. Required for vesicular recycling of CDH1 and TF. May also function in lipid trafficking. Protects cells from spindle disruption damage. Functions in p53/TP53-dependent mitotic spindle checkpoint. Regulates microtubule dynamics and maintains euploidy.

Alternative names

CAP43, DRG1, RTP, NDRG1, Protein NDRG1, Differentiation-related gene 1 protein, N-myc downstream-regulated gene 1 protein, Nickel-specific induction protein Cap43, Reducing agents and tunicamycin-responsive protein, Rit42, DRG-1

Recommended products

Rabbit Recombinant Monoclonal NDRG1 antibody. Suitable for IHC-P, IP, Flow Cyt (Intra), WB, ICC/IF and reacts with Human, Mouse, Rat samples. Cited in 27 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR5593
Purification technique: Affinity purification Protein A
Specificity: PBS only lot tested.
Dissociation constant: 1.33 x 10^-10 M
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

NDRG1 or N-Myc downstream-regulated gene 1 also known as Cap43 Drg-1 and RTP is a protein that plays key roles in various cellular processes. It has a molecular mass of approximately 43 kDa. NDRG1 is ubiquitously expressed with higher levels in the brain heart liver and kidney. The protein also exhibits a significant presence in cytoplasm but can translocate to the nucleus depending on the cellular context and stimuli.

Biological function summary

NDRG1 functions in cell growth regulation stress response and apoptosis influencing differentiation and potentially acting as a tumor suppressor. It does not directly form part of a larger protein complex but interacts with other proteins to mediate its effects. Its involvement in cellular homeostasis places it as a significant player in maintaining normal cell physiology.

Pathways

NDRG1 is involved in the hypoxia and stress response pathways regulating cellular responses to low oxygen and oxidative stress. It interacts with HIF-1α a critical regulator of the cellular response to hypoxia and also modulates PTEN and p53 proteins that are important for cell cycle control and apoptosis. These pathways and interactions reflect its function in cellular regulatory networks.

Associated diseases and disorders

NDRG1 has associations with cancer and Charcot-Marie-Tooth disease type 4D. In cancer abnormal expression of NDRG1 has been linked to tumor progression and metastasis where it interacts with proteins like E-cadherin to influence cell adhesion and migration. In Charcot-Marie-Tooth disease mutations in the NDRG1 gene disrupt normal nerve function pointing to its importance in peripheral nervous system health.

Product promise

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16 product images

Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689)
Lanes 1-4: Merged signal (red and green). Green - ab124689 observed at 43 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
ab124689 Anti-NDRG1 antibody [EPR5593] was shown to specifically react with NDRG1 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human NDRG1 knockout HEK-293T cell line ab267301 (knockout cell lysate Human NDRG1 knockout HEK-293T cell lysate ab257551) was used. Wild-type and NDRG1 knockout samples were subjected to SDS-PAGE. ab124689 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689) at 1/1000 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: NDRG1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: Western blot - Human NDRG1 knockout HEK-293T cell line (Human NDRG1 knockout HEK-293T cell line ab267301)
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDa
Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689)
Lanes 1 - 4: Merged signal (red and green). Green - ab124689 observed at 43 kDa. Red - loading control, Anti-Vinculin antibody [VIN-54] ab130007, observed at 130 kDa.
ab124689 was shown to recognize in wild-type HEK-293 cells as signal was lost at the expected MW in NDRG1 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and NDRG1 knockout samples were subjected to SDS-PAGE. ab124689 and Anti-Vinculin antibody [VIN-54] ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Lanes 1 - 4: Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689) at 1/10000 dilution
Lane 1: Western blot - Alexa Fluor® 647 Anti-NDRG1 antibody [EPR5593] (Alexa Fluor® 647 Anti-NDRG1 antibody [EPR5593] ab199471)
Lane 1: Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2: NDRG1 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 2: Western blot - Human NDRG1 knockout HEK-293 cell line (Human NDRG1 knockout HEK-293 cell line ab261856)
Lane 3: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Predicted band size: 43 kDa
Immunoprecipitation - Anti-NDRG1 antibody [EPR5593] (ab124689)
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg.

Lane 2 (+): ab124689 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab124689 in HeLa whole cell lysate

ab124689 (Purified) at 1/50 dilution (20μg/ml) immunoprecipitating NDRG1 in HeLa whole cell lysate. For western blotting, ab124689 at 1/500 dilution (1.86 μg/mL) and VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1/1000 dilution.

Blocking and diluting buffer: 5% NFDM /TBST .
All lanes: Immunoprecipitation - Anti-NDRG1 antibody [EPR5593] (ab124689)
Predicted band size: 43 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDRG1 antibody [EPR5593] (ab124689)
ab124689 at 1/100 dilution, staining NDRG1 in Formalin fixed Paraffin-embedded human colon tissue by Immunohistochemistry.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-NDRG1 antibody [EPR5593] (ab124689)
ab124689, at 1/100 dilution, staining NDRG1 in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDRG1 antibody [EPR5593] (ab124689)
ab124689 showing positive staining in human normal kidney tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDRG1 antibody [EPR5593] (ab124689)
Immunohistochemical analysis of Paraffin-embedded sections human liver carcinoma tissue labelling NDRG1 with ab124689 at 1/1000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on human liver carcinoma tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.
All lanes: Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689) at 1/10000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: Mouse brain lysate at 20 µg
Lane 3: Rat brain lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 43 kDa
Observed band size: 48 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDRG1 antibody [EPR5593] (ab124689)
Immunohistochemical analysis of Paraffin-embedded sections human colon tissue labelling NDRG1 with ab124689 at 1/1000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on human colon tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689)
NDRG1 Western blot staining using rabbit Anti-NDRG1 antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Insulin treatment increased the expression of phospho T346 NDRG1 (PMID: 18787837).

In Western blot, anti-NDRG1 antibody (ab124689) Pan control staining at 1/1000 dilution.

In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) loading control staining at 1/10000 dilution.

Exposure time: Lane 1 and 2: 70 seconds, Lane 3 and 4: 48 seconds.
All lanes: Western blot - Anti-NDRG1 (phospho T346) antibody [EPR26325-247] (Anti-NDRG1 (phospho T346) antibody [EPR26325-247] ab307706) at 1/1000 dilution
Lane 1: C2C12 whole cell lysate at 20 µg
Lane 2: C2C12 treated with 20nM insulin, whole cell lysate at 20 µg
Lane 3: Rat-1 starved for 4 hours, whole cell lysate at 20 µg
Lane 4: Rat-1 starved for 4 hours, then treated with 20nM insulin, whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/5000 dilution
Observed band size: 48 kDa
Exposure time: 70s
Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

LY294002 treatment decreased the expression of phospho T346 NDRG1(PMID: 18787837).

In Western blot, anti-NDRG1 antibody (ab124689) Pan control staining at 1/1000 dilution.

In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.

Alkaline phosphatase was used to treated membrane.

Exposure time: 103 seconds.
All lanes: Western blot - Anti-NDRG1 (phospho T346) antibody [EPR26325-247] (Anti-NDRG1 (phospho T346) antibody [EPR26325-247] ab307706) at 1/1000 dilution
Lane 1: HeLa starved for 4 hours, whole cell lysate at 20 µg
Lane 2: HeLa starved for 4 hours, then treated with 50uM LY294002 for 4 hours, whole cell lysate at 20 µg
Lane 3: HeLa starved for 4 hours, whole cell lysate (phosphatase treated membrane)
Lane 4: HeLa starved for 4 hours, then treated with 50uM LY294002 for 4 hours, whole cell lysate (phosphatase treated membrane)
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/5000 dilution
Observed band size: 48 kDa
Exposure time: 103s
Immunocytochemistry/ Immunofluorescence - Anti-NDRG1 antibody [EPR5593] (ab124689)
Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling NDRG1 using ab124689. The cells were fixed with 100% Methanol then permeabilized with 0.1% Triton X-100. The cells were then incubated with ab124689 at 1:50 dilution followed by a further incubation with a Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI. Cells were counterstained using Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 dilution (shown in red). Secondary antibody only control: PBS instead of the primary antibody.
Flow Cytometry (Intracellular) - Anti-NDRG1 antibody [EPR5593] (ab124689)
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling NDRG1 with purified ab124689 at 1/20 dilution (5 ug/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as a isotype control. Cell without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDRG1 antibody [EPR5593] (ab124689)
Immunohistochemical analysis of Paraffin-embedded sections mouse colon tissue labelling NDRG1 with ab124689 at 1/1000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on mouse colon tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NDRG1 antibody [EPR5593] (ab124689)
Immunohistochemical analysis of Paraffin-embedded sections rat colon tissue labelling NDRG1 with ab124689 at 1/1000 dilution, followed by a ready to use secondary Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101). Staining on rat colon tissue is observed. Counter stained with Haematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use Rabbit specific IHC polymer detection kit HRP/DAB (Rabbit specific IHC polymer detection kit HRP/DAB ab209101).

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-NDRG1 antibody [EPR5593] (ab124689)
NDRG1 western blot using anti-NDRG1 antibody [EPR5593] ab124689. Publication image and figure legend from Tang, M. K., Liang, Y. J., et al., 2013, PLoS One, PubMed 23555679.

ab124689 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab124689 please see the product overview.
Silencing NDRG1 in PML+/+ MEFs alters gene expression.Semi-quantitative RT-PCR analysis revealed that silencing NDRG1 inhibited Ndrg1, Pml and p53 expression. β-actin served as an internal control for normalization.