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Product Info Summary
| SKU: | M00129-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse, Rat |
| Host: | Mouse |
| Application: | Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-IDH1 Antibody Picoband® (monoclonal, 16H7)
SKU/Catalog Number
M00129-1
Size
100 μg/vial
Form
Liquid
Description
Boster Bio Anti-IDH1 Antibody Picoband® (monoclonal, 16H7) catalog # M00129-1. Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
Storage & Handling
Store at -20˚C for one year from date of receipt. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for six months. Avoid repeated freeze-thaw cycles.
Cite This Product
Anti-IDH1 Antibody Picoband® (monoclonal, 16H7) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00129-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
16H7
Isotype
Mouse IgG1
Immunogen
A synthetic peptide corresponding to a sequence at the C-terminus of human IDH1, different from the related mouse and rat sequences by one amino acid.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00129-1 is reactive to IDH1 in Human, Mouse, Rat
Observed Molecular Weight
47 kDa
Calculated molecular weight
46.7 kDa
Background of IDH1
Isocitrate dehydrogenase 1 (NADP+), soluble is an enzyme that in humans is encoded by the IDH1 gene. Isocitrate dehydrogenases catalyze the oxidative decarboxylation of isocitrate to 2-oxoglutarate. These enzymes belong to two distinct subclasses, one of which utilizes NAD (+) as the electron acceptor and the other NADP (+). Five isocitrate dehydrogenases have been reported: three NAD (+)-dependent isocitrate dehydrogenases, which localize to the mitochondrial matrix, and two NADP (+)-dependent isocitrate dehydrogenases, one of which is mitochondrial and the other predominantly cytosolic. Each NADP (+)-dependent isozyme is a homodimer. The protein encoded by this gene is the NADP (+)-dependent isocitrate dehydrogenase found in the cytoplasm and peroxisomes. It contains the PTS-1 peroxisomal targeting signal sequence. The presence of this enzyme in peroxisomes suggests roles in the regeneration of NADPH for intraperoxisomal reductions, such as the conversion of 2, 4-dienoyl-CoAs to 3-enoyl-CoAs, as well as in peroxisomal reactions that consume 2-oxoglutarate, namely the alpha-hydroxylation of phytanic acid. The cytoplasmic enzyme serves a significant role in cytoplasmic NADPH production. Alternatively spliced transcript variants encoding the same protein have been found for this gene.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
Application & Images
Applications
M00129-1 is guaranteed for Flow Cytometry, IF, ICC, WB Boster Guarantee
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.1-0.5μg/ml, Human, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human HepG2 tissue, human Caco-2 whole cell, human U-87MG whole cell, human THP-1 whole cell, human Hela whole cellhuman K562 whole cellhuman PC-3 whole cellhuman HEK293 whole cell, rat liver tissue, rat RH35 whole cell, mouse liver whole cell,
ICC/IF: U2OS cell
FCM: CACO-2 cell
Validation Images & Assay Conditions
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Western blot analysis of IDH1 using anti-IDH1 antibody (M00129-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: human HepG2 tissue lysates,
Lane 2: human Caco-2 whole cell lysates,
Lane 3: human U-87MG whole cell lysates,
Lane 4: human THP-1 whole cell lysates,
Lane 5: human Hela whole cell lysates.
Lane 6: human K562 whole cell lysates.
Lane 7: human PC-3 whole cell lysates.
Lane 8: human HEK293 whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-IDH1antigen affinity purified polyclonal antibody (Catalog # M00129-1) at 0.5 μg/mL overnight at 4℃, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for IDH1 at approximately 47KD. The expected band size for IDH1 is at 47KD.
Click image to see more details
Western blot analysis of IDH1 using anti-IDH1 antibody (M00129-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: rat RH35 whole cell lysates,
Lane 3: mouse liver whole cell lysates,
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-IDH1antigen affinity purified polyclonal antibody (Catalog # M00129-1) at 0.5 μg/mL overnight at 4℃, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for IDH1 at approximately 47KD. The expected band size for IDH1 is at 47KD.
Click image to see more details
IF analysis of IDH1 using anti-IDH1 antibody (M00129-1).
IDH1 was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-IDH1 Antibody (M00129-1) overnight at 4℃. DyLight488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37℃. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
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Flow Cytometry analysis of CACO-2 cells using anti-IDH1 antibody (M00129-1).
Overlay histogram showing CACO-2 cells stained with M00129-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-IDH1 Antibody (M00129-1, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
Specific Publications For Anti-IDH1 Antibody Picoband® (monoclonal, 16H7) (M00129-1)
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5 Customer Q&As for Anti-IDH1 Antibody Picoband® (monoclonal, 16H7)
Question
My team were happy with the WB result of your anti-IDH1 antibody (monoclonal, 16H7). However we have been able to see positive staining in lung placenta cytoplasm using this antibody. Is that expected? Could you tell me where is IDH1 supposed to be expressed?
Verified Customer
Verified customer
Asked: 2019-12-06
Answer
According to literature, lung placenta does express IDH1. Generally IDH1 expresses in cytoplasm, cytosol. Regarding which tissues have IDH1 expression, here are a few articles citing expression in various tissues:
Brain, Cajal-Retzius cell, and Fetal brain cortex, Pubmed ID: 19608861
Endometrium, Pubmed ID: 17974005
Kidney, Pubmed ID: 11230166
Liver, Pubmed ID: 24275569
Lung, and Placenta, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2019-12-06
Question
We are interested in using your anti-IDH1 antibody (monoclonal, 16H7) for glyoxylate cycle studies. Has this antibody been tested with western blotting on a431 cell lysate? We would like to see some validation images before ordering.
Verified Customer
Verified customer
Asked: 2018-12-19
Answer
Thank you for your inquiry. This M00129-1 anti-IDH1 antibody (monoclonal, 16H7) is validated on a431 cell lysate. It is guaranteed to work for Flow Cytometry, IF, ICC, WB in human, mouse, rat. Our Boster guarantee will cover your intended experiment even if the sample type has not been be directly tested.
Boster Scientific Support
Answered: 2018-12-19
Question
We purchased anti-IDH1 antibody (monoclonal, 16H7) for WB on brain a few years ago. I am using human, and We intend to use the antibody for Flow Cytometry next. I would like examining brain as well as cajal-retzius cell fetal brain cortex in our next experiment. Could you please give me some suggestion on which antibody would work the best for Flow Cytometry?
Verified Customer
Verified customer
Asked: 2018-10-05
Answer
I have checked the website and datasheets of our anti-IDH1 antibody (monoclonal, 16H7) and I see that M00129-1 has been validated on human in both WB and Flow Cytometry. Thus M00129-1 should work for your application. Our Boster satisfaction guarantee will cover this product for Flow Cytometry in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for Flow Cytometry detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2018-10-05
Question
We are currently using anti-IDH1 antibody (monoclonal, 16H7) M00129-1 for mouse tissue, and we are happy with the Flow Cytometry results. The species of reactivity given in the datasheet says human, mouse, rat. Is it true that the antibody can work on horse tissues as well?
R. Carter
Verified customer
Asked: 2018-01-22
Answer
The anti-IDH1 antibody (monoclonal, 16H7) (M00129-1) has not been tested for cross reactivity specifically with horse tissues, though there is a good chance of cross reactivity. We have an innovator award program that if you test this antibody and show it works in horse you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2018-01-22
Question
We have been able to see staining in mouse metanephros. Any tips? Is anti-IDH1 antibody (monoclonal, 16H7) supposed to stain metanephros positively?
K. Bhatt
Verified customer
Asked: 2016-02-16
Answer
According to literature metanephros does express IDH1. According to Uniprot.org, IDH1 is expressed in metanephros, kidney, endometrium, lung placenta, brain, cajal-retzius cell fetal brain cortex, liver, among other tissues. Regarding which tissues have IDH1 expression, here are a few articles citing expression in various tissues:
Brain, Cajal-Retzius cell, and Fetal brain cortex, Pubmed ID: 19608861
Endometrium, Pubmed ID: 17974005
Kidney, Pubmed ID: 11230166
Liver, Pubmed ID: 24275569
Lung, and Placenta, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2016-02-16
