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Product Info Summary
| SKU: | A01774-1 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human, Mouse |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IF, IHC, ICC, WB |
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Product info
Product Name
Anti-MID1 Antibody Picoband®
SKU/Catalog Number
A01774-1
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-MID1 Antibody Picoband® catalog # A01774-1. Tested in ELISA, Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Mouse. 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-MID1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01774-1)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human MID1 recombinant protein (Position: N226-Q278).
*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
A01774-1 is reactive to MID1 in Human, Mouse
Reconstitution
Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Observed Molecular Weight
75 kDa
Calculated molecular weight
35864 MW
Background of MID1
Midline-1 is a protein found in humans that is encoded by the MID1 gene. The protein encoded by this gene is a member of the tripartite motif (TRIM) family, also known as the 'RING-B box-coiled coil' (RBCC) subgroup of RING finger proteins. The TRIM motif includes three zinc-binding domains, a RING, a B-box type 1 and a B-box type 2, and a coiled-coil region. This protein forms homodimers which associate with microtubules in the cytoplasm. The protein is likely involved in the formation of multiprotein structures acting as anchor points to microtubules. Mutations in this gene have been associated with the X-linked form of Opitz syndrome, which is characterized by midline abnormalities such as cleft lip, laryngeal cleft, heart defects, hypospadias, and agenesis of the corpus callosum. This gene was also the first example of a gene subject to X inactivation in human while escaping it in mouse. Alternative promoter use, alternative splicing and alternative polyadenylation result in multiple transcript variants that have different tissue specificities.
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
A01774-1 is guaranteed for ELISA, 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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human HEK29 whole cell, human Hela whole cell,
IHC: human placenta tissue, mouse brain tissue
ICC/IF: U20S cell
FCM: A431 cell
Validation Images & Assay Conditions
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Figure 1. Western blot analysis of MID1 using anti-MID1 antibody (A01774-1).
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 HEK29 whole cell lysates
Lane 2: human Hela 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 rabbit anti-MID1 antigen affinity purified polyclonal antibody (Catalog # A01774-1) 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-rabbit 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 # EK1002) with Tanon 5200 system. A specific band was detected for MID1 at approximately 75KD. The expected band size for MID1 is at 75KD.
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Figure 2. IHC analysis of MID1 using anti-MID1 antibody (A01774-1).
MID1 was detected in paraffin-embedded section of human placenta 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-MID1 Antibody (A01774-1) 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.
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Figure 3. IHC analysis of MID1 using anti-MID1 antibody (A01774-1).
MID1 was detected in paraffin-embedded section of mouse brain 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-MID1 Antibody (A01774-1) 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.
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Figure 4. IF analysis of MID1 using anti-MID1 antibody (A01774-1).
MID1 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 2μg/mL rabbit anti-MID1 Antibody (A01774-1) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) 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.
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Figure 5. Flow Cytometry analysis of A431 cells using anti-MID1 antibody (A01774-1).
Overlay histogram showing A431 cells stained with A01774-1 (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 rabbit anti-MID1 Antibody (A01774-1,1μg/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 MID1 (Source: Uniprot.org, NCBI)
Gene Name
MID1
Full Name
E3 ubiquitin-protein ligase Midline-1
Weight
35864 MW
Superfamily
TRIM/RBCC family
Alternative Names
E3 ubiquitin-protein ligase Midline-1; Midin; Putative transcription factor XPRF; RING finger protein 59; RING finger protein Midline-1; RING-type E3 ubiquitin transferase Midline-1; Tripartite motif-containing protein 18; MID1; FXY; RNF59; TRIM18; XPRF MID1 BBBG1, FXY, GBBB1, MIDIN, OGS1, OS, OSX, RNF59, TRIM18, XPRF, ZNFXY midline 1 E3 ubiquitin-protein ligase Midline-1|Opitz/BBB syndrome|RING finger protein 59|RING finger protein Midline-1|RING-type E3 ubiquitin transferase Midline-1|midline 1 RING finger protein|putative transcription factor XPRF|tripartite motif protein TRIM18|tripartite motif-containing protein 18|zinc finger on X and Y, mouse, homolog of
*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 MID1, check out the MID1 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 MID1: 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-MID1 Antibody Picoband® (A01774-1)
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Customer Q&As
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1 Customer Q&As for Anti-MID1 Antibody Picoband®
Question
We are currently using anti-MID1 antibody A01774-1 for mouse tissue, and we are content with the ELISA results. The species of reactivity given in the datasheet says human, mouse. Is it likely that the antibody can work on feline tissues as well?
A. Lewis
Verified customer
Asked: 2014-12-15
Answer
The anti-MID1 antibody (A01774-1) has not been tested for cross reactivity specifically with feline 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 feline you can get your next antibody for free. Please contact me if I can help you with anything.
Boster Scientific Support
Answered: 2014-12-15
