Anti-Vinculin VCL Antibody (Monoclonal, hVIN-1)

Product Name

Anti-Vinculin VCL Antibody (Monoclonal, hVIN-1)

View all Vinculin Antibodies

SKU/Catalog Number

MA1103

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-Vinculin VCL Antibody (Monoclonal, hVIN-1) catalog # MA1103. Tested in IF, IHC-F, ICC, WB applications. This antibody reacts with Chicken, Human, Monkey, 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-Vinculin VCL Antibody (Monoclonal, hVIN-1) (Boster Biological Technology, Pleasanton CA, USA, Catalog # MA1103)

Host

Mouse

Contents

Mouse ascites fluid, 1.2% sodium acetate, 2mg BSA, with 0.01mg NaN3 as preservative.

Clonality

Monoclonal

Clone Number

hVIN-1

Isotype

Mouse IgG1

Immunogen

Human vinculin, purified from uterus.

*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

MA1103 is reactive to VCL in Chicken, Human, Monkey, Mouse, Rat

Applications

MA1103 is guaranteed for IF, IHC-F, ICC, WB Boster Guarantee

Observed Molecular Weight

124 kDa

Calculated molecular weight

123.799kDa

Background of Vinculin

Vinculin is a cytoskeletal protein associated with the cytoplasmic face of both cell-cell and cell-extracellular matrix adherens-type junctions, where it is thought to function as one of several interacting proteins involved in anchoring F-actin to the membrane. Both human and chicken embryo sequences of vinculin contain 1,066 amino acids and, furthermore, that the 2 proteins exhibit a high level of sequence identity (greater than 95%). Vinculinis mapped to 10q22.1-q23.

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 1ml of PBS buffer will yield a concentration of 100ug/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, 1-2μg/ml, Human, chicken, monkey, mouse, rat
Immunohistochemistry (Frozen Section), 2-4μg/ml, Human, chicken, mouse, rat, -
Immunocytochemistry/Immunofluorescence, 2μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. IHC analysis of Vinculin using anti-Vinculin antibody (MA1103).
Vinculin was detected in frozen section of human placenta tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Vinculin Antibody (MA1103) 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 2. IHC analysis of Vinculin using anti-Vinculin antibody (MA1103).
Vinculin was detected in frozen section of human placenta tissues. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Vinculin Antibody (MA1103) 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. Western blot analysis of Vinculin using anti-Vinculin antibody (MA1103).
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 HepG2 whole cell lysates,
Lane 3: monkey COS-7 whole cell lysates,
Lane 4: human U-87MG whole cell lysates,
Lane 5: rat heart tissue lysates,
Lane 6: rat PC-12 whole cell lysates,
Lane 7: mouse heart tissue lysates,
Lane 8: mosue NIH/3T3 whole cell 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 mouse anti-Vinculin antigen affinity purified monoclonal antibody (Catalog # MA1103) at 1 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 Vinculin at approximately 124 kDa. The expected band size for Vinculin is at 124 kDa.

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Figure 4. IF analysis of Vinculin using anti-Vinculin antibody (MA1103).
Vinculin was detected in immunocytochemical section of human mammary cancer cell. 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 mouse anti-Vinculin Antibody (MA1103) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) 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.

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

Alternative Names

CMD1W; CMH15; metavinculin; MVCL; VCL; Vinculin VCL CMD1W, CMH15, HEL114, MV, MVCL vinculin vinculin|epididymis luminal protein 114|epididymis secretory sperm binding protein|meta-vinculin|metavinculin|vinculin isoform meta-VCL

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