Anti-HES-5 Antibody

HES-5 is a mammalian basic helix-loop-helix factor that has a distant sequence homology to the product of the Drosophila pair-rule gene hairy. HES-5 mRNA is present exclusively in the developing nervous system, but its level decreases as neural differentiation proceeds.

Synthetic peptide, amino acids 83-95 of mouse HES-5.

Anti-HES-5 Antibody detects level of HES-5 & has been published & validated for use in WB, IH.

Research Category
Neuroscience

Research Sub Category
Developmental Neuroscience

Neuronal & Glial Markers

Western blot: 1:200-1:1,000 using ECL. Detects a 28kDa band in embryonal RN46a rat cell line {http://www.jneurosci.org/cgi/reprint/23/25/8788.pdf}



Immunohistochemistry: 1:200-1:1,000.

Optimal working dilutions must be determined by the end user.

Recognizes HES-5 (Hairy family), a Helix-loop-helix class of transcription factor.

Affinity purified immunoglobulin. Precipitated antibody in a solution of 50% saturated ammonium sulfate and PBS containing no preservatives.

Maintain unopened vial at -70°C for up to 6 months. Avoid repeated freeze/thaw cycles.

PREPARATION AND USE:

To reconstitute the antibody, centrifuge the antibody vial at moderate speed (5,000 rpm) for 5 minutes to pellet the precipitated antibody product. Carefully remove the ammonium sulfate/PBS buffer solution and discard. It is not necessary to remove all of the ammonium sulfate/PBS solution: 10 μL of residual ammonium sulfate solution will not affect the resuspension of the antibody. Do not let the protein pellet dry, as severe loss of antibody reactivity can occur.

Resuspend the antibody pellet in a suitable biological buffer such as PBS or TBS (pH 7.3-7.5) to a final concentration of 1.0 mg/mL. For example, to achieve a 1.0 mg/mL concentration with 50 μg of precipitated antibody, the amount of buffer needed would be 50 μL.

Carefully add the liquid buffer to the pellet. DO NOT VORTEX. Mix by gentle stirring with a wide pipet tip or gentle finger-tapping. Let the precipitated antibody rehydrate for 1 hour at 4-25°C prior to use. Small particles of precipitated antibody that fail to resuspend are normal. Vials are overfilled to compensate for any losses.

The rehydrated antibody solutions can be stored undiluted at 2-8°C for 2 months without any significant loss of activity. Note, the solution is not sterile, thus care should be taken if product is stored at 2-8°C.

For storage at -20°C, the addition of an equal volume of glycerol can be used, however, it is recommended that ACS grade or higher glycerol be used, as significant loss of activity can occur if the glycerol used is not of high quality.

For long-term storage at -70°C, it is recommended that the rehydrated antibody solution be further diluted 1:1 with a 2% BSA (fraction V, highest-grade available) solution made with the rehydration buffer. The resulting 1% BSA/antibody solution can be aliquoted and stored frozen at -70°C for up to 6 months. Avoid repeated freeze/thaw cycles.

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