Anti-NDRG1 antibody [EPR5593] - Low endotoxin, Azide free (ab216457)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5593] to NDRG1 - Low endotoxin, Azide free
- Suitable for: ICC, IP, IHC-P, WB
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-NDRG1 antibody [EPR5593] - Low endotoxin, Azide free
See all NDRG1 primary antibodies -
Description
Rabbit monoclonal [EPR5593] to NDRG1 - Low endotoxin, Azide free -
Host species
Rabbit -
Tested applications
Suitable for: ICC, IP, IHC-P, WBmore details
Unsuitable for: Flow Cyt -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat -
Immunogen
This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human colon tissue, cervical carcinoma tissue, colonic adenocarcinoma tissue, normal kidney tissue and breast carcinoma tissue. ICC/IF: HeLa. IP: HeLa. WB: Wild-type HEK-293 whole cell lysate. Jurkat, HeLa, Caco-2 and LnCap whole cell lysate. Mouse and rat brain lysate.
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General notes
ab216457 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Dissociation constant (KD)
KD = 1.33 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR5593 -
Isotype
IgG -
Research areas
Images
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All lanes : 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
Lane 2 : NDRG1 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 43 kDaThis data was developed using ab124689, the same antibody clone in a different buffer formulation.
Lanes 1-4: Merged signal (red and green). Green - ab124689 observed at 43 kDa. Red - 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 ab267301 (knockout cell lysate ab257551) was used. Wild-type and NDRG1 knockout samples were subjected to SDS-PAGE. ab124689 and 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 (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.
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This IHC data was generated using the same anti-NDRG1 antibody clone, EPR5593, in a different buffer formulation (cat# ab124689).
ab124689 at 1/100 dilution, staining NDRG1 in Formalin fixed Paraffin-embedded Human colon tissue by Immunohistochemistry.
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-NDRG1 antibody [EPR5593] (ab124689) at 1/10000 dilution
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 : NDRG1 knockout HEK-293 whole cell lysate
Lane 3 : Jurkat whole cell lysate
Lane 4 : LnCap whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 43 kDaThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
Lanes 1 - 4: Merged signal (red and green). Green - ab124689 observed at 43 kDa. Red - loading control, 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 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 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.
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ab124689 showing positive staining in Cervical carcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Clone EPR5593 (ab216457) has been successfully conjugated by Abcam. This image was generated using Anti-NDRG1 antibody [EPR5593] (Alexa Fluor® 647). Please refer to ab199471 for protocol details.
ab199471 staining NDRG1 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab at a 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at a 1/250 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Clone EPR5593 (ab216457) has been successfully conjugated by Abcam. This image was generated using Anti-NDRG1 antibody [EPR5593] (Alexa Fluor® 488). Please refer to ab199233 for protocol details.
ab199233 staining NDRG1 in HeLa cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab199233 at 1/500 dilution (shown in green) and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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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 (ab172730) instead of ab124689 in HeLa whole cell lysate
Ab124689 (Purified) at 1:500 dilution (1.86 µg/ml) immunoprecipitating NDRG1 in HeLa whole cell lysate. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1,000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST .This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
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ab124689 showing positive staining in Colonic adenocarcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab124689 showing positive staining in Normal kidney tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab124689 showing positive staining in Breast carcinoma tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124689).
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