Anti-ARAF Antibody Picoband™

Product Name

Anti-ARAF Antibody Picoband™

View all A-RAF Antibodies

SKU/Catalog Number

A02061-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-ARAF Antibody Picoband™ catalog # A02061-2. Tested in ELISA, Flow Cytometry, IF, 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-ARAF Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A02061-2)

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 ARAF recombinant protein (Position: E309-T413).

*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

A02061-2 is reactive to ARAF in Human, Mouse, Rat

Applications

A02061-2 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee

Observed Molecular Weight

68 kDa

Calculated molecular weight

67.585kDa

Background of A-RAF

Serine/threonine-protein kinase A-Raf or simply A-Raf is an enzyme that in humans is encoded by the ARAF gene. It is mapped to Xp11.3. This proto-oncogene belongs to the RAF subfamily of the Ser/Thr protein kinase family, and maybe involved in cell growth and development. Alternatively spliced transcript variants encoding different isoforms have been found for this gene. A-Raf is a member of the Raf kinase family of serine/threonine-specific protein kinases.

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, Mouse, Rat
Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
Flow Cytometry, 1-3 μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of ARAF using anti-ARAF antibody (A02061-2).
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 MCF-7 whole cell lysates,
Lane 3: rat testis tissue lysates,
Lane 4: rat brain tissue lysates,
Lane 5: mouse testis tissue lysates,
Lane 6: mouse brain 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-ARAF antigen affinity purified polyclonal antibody (Catalog # A02061-2) 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 ARAF at approximately 68 kDa. The expected band size for ARAF is at 68 kDa.

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Figure 2. IF analysis of ARAF using anti-ARAF antibody (A02061-2).
ARAF was detected in an immunocytochemical section of MCF-7 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-ARAF Antibody (A02061-2) 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.

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Figure 3. IF analysis of ARAF using anti-ARAF antibody (A02061-2).
ARAF was detected in an immunocytochemical section of PC-3 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-ARAF Antibody (A02061-2) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) 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 4. Flow Cytometry analysis of SiHa cells using anti-ARAF antibody (A02061-2).
Overlay histogram showing SiHa cells stained with A02061-2 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ARAF Antibody (A02061-2, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

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

Alternative Names

ARaf; A-Raf; ARAF1A-RAF; EC 2.7.11; EC 2.7.11.1; Oncogene ARAF1; PKS; PKS2; PKS2A-Raf proto-oncogene serine/threonine-protein kinase; Proto-oncogene A-Raf; Proto-oncogene A-Raf-1; Proto-oncogene Pks; RAFA1; Ras-binding protein DA-Raf; serine/threonine-protein kinase A-Raf; v-raf murine sarcoma 3611 viral oncogene homolog 1; v-raf murine sarcoma 3611 viral oncogene homolog ARAF A-RAF1, PKS2, RAFA1, ARAF A-Raf proto-oncogene, serine/threonine kinase serine/threonine-protein kinase A-Raf|A-Raf proto-oncogene serine/threonine-protein kinase|Oncogene ARAF1|Ras-binding protein DA-Raf|proto-oncogene A-Raf-1|proto-oncogene Pks|v-raf murine sarcoma 3611 viral oncogene homolog 1|v-raf murine sarcoma 3611 viral oncogene-like protein

For more info on ARAF, check out the ARAF Infographic

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