Product Info Summary
SKU: | PB10039 |
---|---|
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-AMFR Antibody Picoband™
SKU/Catalog Number
PB10039
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-AMFR Antibody Picoband™ catalog # PB10039. Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
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-AMFR Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB10039)
Host
Rabbit
Contents
Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E. coli-derived human AMFR recombinant protein (Position: E553-S643). Human AMFR shares 89% amino acid (aa) sequence identity with mouse AMFR.
*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
PB10039 is reactive to AMFR in Human, Mouse, Rat
Applications
PB10039 is guaranteed for Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee
Observed Molecular Weight
73 kDa
Calculated molecular weight
72.996kDa
Background of AMFR/gp78
Autocrine motility factor receptor, isoform 2 is a protein that in humans is encoded by the AMFR gene. Autocrine motility factor is a tumor motility-stimulating protein secreted by tumor cells. The protein encoded by this gene is a glycosylated transmembrane protein and a receptor for autocrine motility factor. The receptor, which shows some sequence similarity to tumor protein p53, is localized to the leading and trailing edges of carcinoma cells. Its ligand, autocrine motility factor, is a tumor motility-stimulating protein secreted by tumor cells. The encoded receptor is also a member of the E3 ubiquitin ligase family of proteins. It catalyzes ubiquitination and endoplasmic reticulum-associated degradation of specific proteins.
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.
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Assay dilution & Images
Reconsitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
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, Rat
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, Rat, By Heat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3μg/1x106 cells, Human
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of AMFR using anti-AMFR antibody (PB10039).
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 HepG2 whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human Daudi whole cell lysates,
Lane 4: rat 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-AMFR antigen affinity purified polyclonal antibody (Catalog # PB10039) 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 AMFR at approximately 73 kDa. The expected band size for AMFR is at 73 kDa.
Click image to see more details
Figure 2. IHC analysis of AMFR using anti-AMFR antibody (PB10039).
AMFR was detected in a paraffin-embedded section of human placenta 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 1 μg/ml rabbit anti-AMFR Antibody (PB10039) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 3. IHC analysis of AMFR using anti-AMFR antibody (PB10039).
AMFR was detected in paraffin-embedded section of rat kidney tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-AMFR Antibody (PB10039) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 4. IHC analysis of AMFR using anti-AMFR antibody (PB10039).
AMFR was detected in paraffin-embedded section of mouse intestine tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-AMFR Antibody (PB10039) overnight at 4°C. Biotinylated goat anti-rabbit 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 # SA1022) with DAB as the chromogen.
Click image to see more details
Figure 5. Flow Cytometry analysis of SiHa cells using anti-AMFR antibody (PB10039).
Overlay histogram showing SiHa cells stained with PB10039 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AMFR Antibody (PB10039,1μg/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 (Red line) was also used as a control.
Click image to see more details
Figure 6. Flow Cytometry analysis of U87 cells using anti-AMFR antibody (PB10039).
Overlay histogram showing SiHa cells stained with PB10039 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-AMFR Antibody (PB10039,1μg/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 (Red line) was also used as a control.
Click image to see more details
Figure 7. IF analysis of AMFR using anti-AMFR antibody (PB10039).
AMFR was detected in an immunocytochemical section of T47D 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 5 μg/mL rabbit anti-AMFR Antibody (PB10039) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 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.
Protein Target Info & Infographic
Gene/Protein Information For AMFR (Source: Uniprot.org, NCBI)
Gene Name
AMFR
Full Name
E3 ubiquitin-protein ligase AMFR
Weight
72.996kDa
Alternative Names
AMF receptor; AMF receptor, isoform 1; AMF receptor, isoform 2; AMFR; autocrine motility factor receptor; Autocrine motility factor receptor, isoform 2; gp78; RING finger protein 45; RNF45; RNF45EC 6.3.2.- AMFR GP78, RNF45 autocrine motility factor receptor E3 ubiquitin-protein ligase AMFR|RING finger protein 45|RING-type E3 ubiquitin transferase AMFR
*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 AMFR, check out the AMFR 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 AMFR: 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-AMFR Antibody Picoband™ (PB10039)
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No publications found for PB10039
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Customer Q&As
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5 Customer Q&As for Anti-AMFR Antibody Picoband™
Question
I am looking for to test anti-AMFR antibody PB10039 on rat liver for research purposes, then I may be interested in using anti-AMFR antibody PB10039 for diagnostic purposes as well. Is the antibody suitable for diagnostic purposes?
Verified Customer
Verified customer
Asked: 2019-08-28
Answer
The products we sell, including anti-AMFR antibody PB10039, 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: 2019-08-28
Question
We are currently using anti-AMFR antibody PB10039 for mouse tissue, and we are satisfied 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 feline tissues as well?
Verified Customer
Verified customer
Asked: 2019-05-27
Answer
The anti-AMFR antibody (PB10039) has not been tested for cross reactivity specifically with feline tissues, but 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 feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2019-05-27
Question
Does PB10039 anti-AMFR antibody work on parafin embedded sections? If so, which fixation method do you recommend we use (PFA, paraformaldehyde, other)?
Verified Customer
Verified customer
Asked: 2019-02-01
Answer
As indicated on the product datasheet, PB10039 anti-AMFR antibody as been tested on Flow Cytometry. 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: 2019-02-01
Question
We appreciate helping with my inquiry over the phone. Here are the WB image, lot number and protocol we used for liver using anti-AMFR antibody PB10039. Let me know if you need anything else.
Verified Customer
Verified customer
Asked: 2018-02-28
Answer
I appreciate 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: 2018-02-28
Question
See attached the WB image, lot number and protocol we used for liver using anti-AMFR antibody PB10039. Please let me know if you require anything else.
T. Johnson
Verified customer
Asked: 2014-01-08
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: 2014-01-08