Anti-MID1 Antibody Picoband™

Product Name

Anti-MID1 Antibody Picoband™

View all MID1 Antibodies

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.

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

Applications

A01774-1 is guaranteed for ELISA, Flow Cytometry, IF, IHC, ICC, WB Boster Guarantee

Observed Molecular Weight

75 kDa

Calculated molecular weight

75.251kDa

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.

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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
Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml, Human, Mouse, By Heat
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

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).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 (Red line) was also used as a control.

Gene/Protein Information For MID1 (Source: Uniprot.org, NCBI)

Alternative Names

EC 6.3.2.-; FLJ57031; FLJ58683; FLJ76288; FXYGBBB1; Midin; midline 1 (Opitz/BBB syndrome); RNF59MIDIN; TRIM18mouse, homolog of 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

For more info on MID1, check out the MID1 Infographic

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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].

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