Product Info Summary
SKU: | PB9175 |
---|---|
Size: | 100 μg/vial |
Reactive Species: | Human, Mouse, Rat |
Host: | Rabbit |
Application: | Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-Dihydrofolate reductase (DHFR) Antibody Picoband®
View all Dihydrofolate Reductase/DHFR Antibodies
SKU/Catalog Number
PB9175
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-Dihydrofolate reductase (DHFR) Antibody Picoband® catalog # PB9175. Tested in Flow Cytometry, IF, IHC, 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-Dihydrofolate reductase (DHFR) Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9175)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.01mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human DHFR recombinant protein (Position: V2-D187). Human DHFR shares 90% amino acid (aa) sequence identity with both mouse and rat 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
PB9175 is reactive to DHFR in Human, Mouse, Rat
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
21 kDa
Calculated molecular weight
21453 MW
Background of Dihydrofolate Reductase/DHFR
Dihydrofolate reductase, or DHFR, is an enzyme that reduces dihydrofolic acid to tetrahydrofolic acid, using NADPH as electron donor, which can be converted to the kinds of tetrahydrofolate cofactors used in 1-carbon transfer chemistry. In humans, the DHFR enzyme is encoded by the DHFR gene. It is found in the q11→q22 region of chromosome 5. What's more, DHFR belongs to the dihydrofolate reductase family, and it converts dihydrofolate into tetrahydrofolate, a methyl group shuttle required for the de novo synthesis of purines, thymidylic acid, and certain amino acids. DHFR is the key enzyme in folate metabolism. In addition, DHFR catalyzes an essential reaction for de novo glycine and purine synthesis, and for DNA precursor synthesis.
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
PB9175 is guaranteed for Flow Cytometry, IF, IHC, 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
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
Positive Control
WB: human Hela whole cell, human Jurkat whole cell, rat liver tissue, rat kidney tissue, mouse liver tissue, mouse kidney tissue
IHC: human colorectal cancer tissue, mouse small intestine tissue, rat small intestine tissue
ICC/IF: U20S cell
FCM: PC-3 cell
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of DHFR using anti-DHFR antibody (PB9175).
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 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: rat liver tissue lysates,
Lane 4: rat kidney tissue lysates,
Lane 5: mouse liver tissue lysates,
Lane 6: mouse kidney tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-DHFR antigen affinity purified polyclonal antibody (Catalog # PB9175) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for DHFR at approximately 21 kDa. The expected band size for DHFR is at 21 kDa.
Click image to see more details
Figure 2. IHC analysis of DHFR using anti-DHFR antibody (PB9175).
DHFR was detected in a paraffin-embedded section of human colorectal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-DHFR Antibody (PB9175) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of DHFR using anti-DHFR antibody (PB9175).
DHFR was detected in a paraffin-embedded section of mouse small intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-DHFR Antibody (PB9175) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of DHFR using anti-DHFR antibody (PB9175).
DHFR was detected in a paraffin-embedded section of rat small intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-DHFR Antibody (PB9175) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.
Click image to see more details
Figure 5. IF analysis of DHFR using anti-DHFR antibody (PB9175).
DHFR was detected in immunocytochemical section of U20S cells. 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 5μg/mL rabbit anti-DHFR Antibody (PB9175) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 6. Flow Cytometry analysis of PC-3 cells using anti-DHFR antibody (PB9175).
Overlay histogram showing PC-3 cells stained with PB9175 (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 rabbit anti-DHFR Antibody (PB9175, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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.
Protein Target Info & Infographic
Gene/Protein Information For DHFR (Source: Uniprot.org, NCBI)
Gene Name
DHFR
Full Name
Dihydrofolate reductase
Weight
21453 MW
Superfamily
dihydrofolate reductase family
Alternative Names
Dihydrofolate reductase;1.5.1.3;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-Dihydrofolate reductase (DHFR) Antibody Picoband® (PB9175)
Hello CJ!
PB9175 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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17 Customer Q&As for Anti-Dihydrofolate reductase (DHFR) Antibody Picoband®
Question
Does PB9175 anti-Dihydrofolate reductase (DHFR) antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2020-05-05
Answer
It shows on the product datasheet, PB9175 anti-Dihydrofolate reductase (DHFR) antibody as been tested on IHC. It is best to use PFA for fixation because it has better tissue penetration ability. PFA needs to be prepared fresh before use. Long term stored PFA turns into formalin, as the PFA molecules congregate and become formalin.
Boster Scientific Support
Answered: 2020-05-05
Question
Can you help my question with product PB9175, anti-Dihydrofolate reductase (DHFR) antibody. I was wondering if it would be possible to conjugate this antibody with biotin. I would need it to be without BSA or sodium azide. I am planning on using a buffer exchange of sodium azide with PBS only. Would there be problems for me to conjugate the antibody and store it in -20 degrees in small aliquots?
Verified Customer
Verified customer
Asked: 2020-04-21
Answer
We do not advise storing this antibody with PBS buffer only in -20 degrees. If you want to store it in -20 degrees it is best to add some cryoprotectant like glycerol. If you want carrier free PB9175 anti-Dihydrofolate reductase (DHFR) antibody, we can provide it to you in a special formula with trehalose and/or glycerol. These molecules will not interfere with conjugation chemistry and provide a good level of protection for the antibody from degradation. Please be sure to specify this in your purchase order.
Boster Scientific Support
Answered: 2020-04-21
Question
Is there a BSA free version of anti-Dihydrofolate reductase (DHFR) antibody PB9175 available?
Verified Customer
Verified customer
Asked: 2020-03-04
Answer
I appreciate your recent telephone inquiry. I can confirm that some lots of this anti-Dihydrofolate reductase (DHFR) antibody PB9175 are BSA free. For now, these lots are available and we can make a BSA free formula for you free of charge. It will take 3 extra days to prepare. If you require this antibody BSA free again in future, please do not hesitate to contact me and I will be pleased to check which lots we have in stock that are BSA free.
Boster Scientific Support
Answered: 2020-03-04
Question
We were well pleased with the WB result of your anti-Dihydrofolate reductase (DHFR) antibody. However we have observed positive staining in buccal mucosa cell mitochondrion using this antibody. Is that expected? Could you tell me where is DHFR supposed to be expressed?
B. Banerjee
Verified customer
Asked: 2019-08-19
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: 2019-08-19
Question
I was wanting to use your anti-Dihydrofolate reductase (DHFR) antibody for IHC for human eye on frozen tissues, but I want to know if it has been tested for this particular application. Has this antibody been tested and is this antibody a good choice for human eye identification?
Verified Customer
Verified customer
Asked: 2019-08-01
Answer
As indicated on the product datasheet, PB9175 anti-Dihydrofolate reductase (DHFR) antibody has been validated for IHC, WB on human, mouse, rat tissues. We have an innovator award program that if you test this antibody and show it works in human eye in IHC-frozen, you can get your next antibody for free.
Boster Scientific Support
Answered: 2019-08-01
Question
I see that the anti-Dihydrofolate reductase (DHFR) antibody PB9175 works with IHC, what is the protocol used to produce the result images on the product page?
Verified Customer
Verified customer
Asked: 2019-06-10
Answer
You can find protocols for IHC on the "support/technical resources" section of our navigation menu. If you have any further questions, please send an email to [email protected]
Boster Scientific Support
Answered: 2019-06-10
Question
Is a blocking peptide available for product anti-Dihydrofolate reductase (DHFR) antibody (PB9175)?
Verified Customer
Verified customer
Asked: 2019-04-12
Answer
We do provide the blocking peptide for product anti-Dihydrofolate reductase (DHFR) antibody (PB9175). If you would like to place an order for it please contact [email protected] and make a special request.
Boster Scientific Support
Answered: 2019-04-12
Question
I appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for eye using anti-Dihydrofolate reductase (DHFR) antibody PB9175. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2018-08-22
Answer
Thank you for the data. You have provided everything we needed. Our lab team are working to resolve your inquiry as quickly as possible, and we appreciate your patience and understanding! Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2018-08-22
Question
We have been able to see staining in human buccal mucosa cell. Any tips? Is anti-Dihydrofolate reductase (DHFR) antibody supposed to stain buccal mucosa cell positively?
Verified Customer
Verified customer
Asked: 2018-08-13
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: 2018-08-13
Question
We need to test anti-Dihydrofolate reductase (DHFR) antibody PB9175 on human eye for research purposes, then I may be interested in using anti-Dihydrofolate reductase (DHFR) antibody PB9175 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2017-11-20
Answer
The products we sell, including anti-Dihydrofolate reductase (DHFR) antibody PB9175, are only intended for research use. They would not be suitable for use in diagnostic work. If you have the means to develop a product into diagnostic use, and are interested in collaborating with us and develop our product into an IVD product, please contact us for more discussions.
Boster Scientific Support
Answered: 2017-11-20
Question
Is this PB9175 anti-Dihydrofolate reductase (DHFR) antibody reactive to the isotypes of DHFR?
Verified Customer
Verified customer
Asked: 2017-09-08
Answer
The immunogen of PB9175 anti-Dihydrofolate reductase (DHFR) antibody is E.coli-derived human DHFR recombinant protein (Position: V2-D187). Human DHFR shares 90% amino acid (aa) sequence identity with both mouse and rat DHFR. Could you tell me which isotype you are interested in so I can help see if the immunogen is part of this isotype?
Boster Scientific Support
Answered: 2017-09-08
Question
Would anti-Dihydrofolate reductase (DHFR) antibody PB9175 work on goat IHC with buccal mucosa cell?
Verified Customer
Verified customer
Asked: 2017-05-15
Answer
Our lab technicians have not tested anti-Dihydrofolate reductase (DHFR) antibody PB9175 on goat. You can run a BLAST between goat and the immunogen sequence of anti-Dihydrofolate reductase (DHFR) antibody PB9175 to see if they may cross-react. If the sequence homology is close, then you can perform a pilot test. Keep in mind that since we have not validated goat samples, this use of the antibody is not covered by our guarantee. However we have an innovator award program that if you test this antibody and show it works in goat buccal mucosa cell in IHC, you can get your next antibody for free.
Boster Scientific Support
Answered: 2017-05-15
Question
We are currently using anti-Dihydrofolate reductase (DHFR) antibody PB9175 for human tissue, and we are satisfied with the WB results. The species of reactivity given in the datasheet says human, mouse, rat. Is it possible that the antibody can work on dog tissues as well?
D. Zhao
Verified customer
Asked: 2017-02-22
Answer
The anti-Dihydrofolate reductase (DHFR) antibody (PB9175) has not been tested for cross reactivity specifically with dog 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 dog you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2017-02-22
Question
Please see the WB image, lot number and protocol we used for eye using anti-Dihydrofolate reductase (DHFR) antibody PB9175. Please let me know if you require anything else.
S. Carter
Verified customer
Asked: 2016-10-28
Answer
Thank you very much for the data. Our lab team are working to resolve this as quickly as possible, and we appreciate your patience and understanding! You have provided everything we needed. Please let me know if there is anything you need in the meantime.
Boster Scientific Support
Answered: 2016-10-28
Question
Would anti-Dihydrofolate reductase (DHFR) antibody PB9175 work for IHC with eye?
S. Li
Verified customer
Asked: 2016-08-01
Answer
According to the expression profile of eye, DHFR is highly expressed in eye. So, it is likely that anti-Dihydrofolate reductase (DHFR) antibody PB9175 will work for IHC with eye.
Boster Scientific Support
Answered: 2016-08-01
Question
Our lab used your anti-Dihydrofolate reductase (DHFR) antibody for WB on eye in the past. I am using human, and We are going to use the antibody for IHC next. We need examining eye as well as buccal mucosa cell in our next experiment. Could you please give me some suggestion on which antibody would work the best for IHC?
G. Dhar
Verified customer
Asked: 2016-03-21
Answer
I took a look at the website and datasheets of our anti-Dihydrofolate reductase (DHFR) antibody and it appears that PB9175 has been validated on human in both WB and IHC. Thus PB9175 should work for your application. Our Boster satisfaction guarantee will cover this product for IHC in human even if the specific tissue type has not been validated. We do have a comprehensive range of products for IHC detection and you can check out our website bosterbio.com to find out more information about them.
Boster Scientific Support
Answered: 2016-03-21
Question
I would like using your anti-Dihydrofolate reductase (DHFR) antibody for recycling studies. Has this antibody been tested with western blotting on tissue lysate? We would like to see some validation images before ordering.
K. Williams
Verified customer
Asked: 2015-07-14
Answer
Thanks for your inquiry. This PB9175 anti-Dihydrofolate reductase (DHFR) antibody is validated on rat liver tissue, tissue lysate, mouse liver tissue, hepg2 whole cell lysate, jurkat whole cell lysate, brain tissue, lung cancer tissue. It is guaranteed to work for IHC, 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: 2015-07-14