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Product Info Summary
| SKU: | M00813-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Rat |
| Host: | Mouse |
| Application: | IHC, WB |
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Product info
Product Name
Anti-DHFR Antibody Picoband® (monoclonal, 3C8)
View all Dihydrofolate Reductase/DHFR Antibodies
SKU/Catalog Number
M00813-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-DHFR Antibody Picoband® (monoclonal, 3C8) catalog # M00813-1. Tested in IHC, WB applications. This antibody reacts with Human, 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-DHFR Antibody Picoband® (monoclonal, 3C8) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M00813-1)
Host
Mouse
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Monoclonal
Clone Number
3C8
Isotype
Mouse IgG2b
Immunogen
E.coli-derived human DHFR recombinant protein (Position: V2-D187). Human DHFRshares90%aminoacid (aa)sequenceidentitywith both mouseandrat DHFR.
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
M00813-1 is reactive to DHFR in Human, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500μg/ml.
Observed Molecular Weight
22 kDa
Calculated molecular weight
21.453kDa
Background of Dihydrofolate Reductase/DHFR
SHC, also known as SHC1 (SHC-transforming protein 1) or SHCA, is a protein that in humans is encoded by the SHC1 gene. SCOP classifies the 3D structure as belonging to the SH2 domain family. This gene encodes three main isoforms that differ in activities and subcellular location. While all three are adapter proteins in signal transduction pathways, the longest (p66Shc) may be involved in regulating life span and the effects of reactive oxygen species. The other two isoforms, p52Shc and p46Shc, link activated receptor tyrosine kinases to the Ras pathway by recruitment of the GRB2/SOS complex. p66Shc is not involved in Ras activation. Unlike the other two isoforms, p46Shc is targeted to the mitochondrial matrix. Several transcript variants encoding different isoforms 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
M00813-1 is guaranteed for IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml
Positive Control
WB: human K562 whole cell, human SGC-7901 whole cell, human Hela whole cell, human SW620 whole cell, rat kidney tissue
IHC: human rectal cancer tissue, rat gaster tissue
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of DHFR using anti-DHFR antibody (M00813-1).
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 K562 whole cell lysates,
Lane 2: human SGC-7901 whole cell lysates,
Lane 3: human Hela whole cell lysates,
Lane 4: human SW620 whole cell lysates,
Lane 5: rat kidney tissue 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-DHFR antigen affinity purified monoclonal antibody (Catalog # M00813-1) at 0.5 μg/mL overnight at 4°C, 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 DHFR at approximately 22KD. The expected band size for DHFR is at 22KD.
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Figure 2. IHC analysis of DHFR using anti-DHFR antibody (M00813-1).
DHFR was detected in paraffin-embedded section of human rectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-DHFR Antibody (M00813-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
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Figure 3. IHC analysis of DHFR using anti-DHFR antibody (M00813-1).
DHFR was detected in paraffin-embedded section of rat gaster tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml mouse anti-DHFR Antibody (M00813-1) overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1021) with DAB as the chromogen.
Protein Target Info & Infographic
Gene/Protein Information For DHFR (Source: Uniprot.org, NCBI)
Gene Name
DHFR
Full Name
Dihydrofolate reductase
Weight
21.453kDa
Superfamily
dihydrofolate reductase family
Alternative Names
Dihydrofolate reductase; DHFR DHFR DHFRP1, DYR dihydrofolate reductase dihydrofolate reductase
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on DHFR, check out the DHFR Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for DHFR: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-DHFR Antibody Picoband® (monoclonal, 3C8) (M00813-1)
Hello CJ!
M00813-1 has been cited in 1 publications:
*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.
Tian L, Zhang J, Ge J, Xiao H, Lu J, Fu S, Liu M, Sun Y. Med Oncol. 2014 Jan;31(1):785. Doi: 10.1007/S12032-013-0785-3. Epub 2013 Dec 3. Microrna-205 Suppresses Proliferation And Promotes Apoptosis In Laryngeal Squamous Cell Carcinoma.
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Customer Q&As
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4 Customer Q&As for Anti-DHFR Antibody Picoband® (monoclonal, 3C8)
Question
We are currently using anti-DHFR antibody (monoclonal, 3C8) M00813-1 for rat tissue, and we are happy with the IHC-P results. The species of reactivity given in the datasheet says human, rat. Is it true that the antibody can work on pig tissues as well?
Verified Customer
Verified customer
Asked: 2019-09-04
Answer
The anti-DHFR antibody (monoclonal, 3C8) (M00813-1) has not been tested for cross reactivity specifically with pig 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 pig you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-09-04
Question
We ordered your anti-DHFR antibody (monoclonal, 3C8) for WB on eye in a previous experiment. I am using rat, and We intend to use the antibody for IHC-P next. We need examining eye as well as buccal mucosa cell in our next experiment. Could give a recommendation on which antibody would work the best for IHC-P?
E. Williams
Verified customer
Asked: 2018-10-03
Answer
I looked at the website and datasheets of our anti-DHFR antibody (monoclonal, 3C8) and I see that M00813-1 has been validated on rat in both WB and IHC-P. Thus M00813-1 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC-P in rat even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC-P detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2018-10-03
Question
My colleagues were satisfied with the WB result of your anti-DHFR antibody (monoclonal, 3C8). However we have been able to see positive staining in buccal mucosa cell mitochondrion using this antibody. Is that expected? Could you tell me where is DHFR supposed to be expressed?
Verified Customer
Verified customer
Asked: 2018-06-26
Answer
According to literature, buccal mucosa cell does express DHFR. Generally DHFR expresses in mitochondrion. Regarding which tissues have DHFR expression, here are a few articles citing expression in various tissues:
Eye, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2018-06-26
Question
We have been able to see staining in rat buccal mucosa cell. Are there any suggestions? Is anti-DHFR antibody (monoclonal, 3C8) supposed to stain buccal mucosa cell positively?
A. Krishna
Verified customer
Asked: 2017-10-31
Answer
According to literature buccal mucosa cell does express DHFR. According to Uniprot.org, DHFR is expressed in buccal mucosa cell, eye, among other tissues. Regarding which tissues have DHFR expression, here are a few articles citing expression in various tissues:
Eye, Pubmed ID: 15489334
Boster Scientific Support
Answered: 2017-10-31
