APEG1 Protein, Human (His)

APEG1 Isoform 3 potentially regulates the growth and differentiation of arterial smooth muscle cells. APEG1 Protein, Human (His) is the recombinant human-derived APEG1 protein, expressed by E. coli , with C-His labeled tag.

APEG1, specifically isoform 3, is implicated in the potential regulation of growth and differentiation processes in arterial smooth muscle cells.

When Recombinant Human APEG1 Protein is immobilized at 10 µg/mL (100 µL/well) can bind Anti- APEG1 Antibody, The ED₅₀ for this effect is 8.941 μg/mL.

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  When Recombinant Human APEG1 Protein is immobilized at 10 µg/mL (100 µL/well) can bind Anti- APEG1 Antibody, The ED₅₀ for this effect is 8.941 μg/mL.

Materials
Biotin
Pre-chilled sterile water
Wash buffer: 1X PBST
Blocking buffer: 1% BSA in 1X PBST
TMB substrate solution: Mix substrate solution A and B at a 1:1 ratio immediately before use
Stop solution: 2 M H₂SO₄
APEG1 Protein, Human (His) (HY-P76729)

Procedure
1. Coating: Dilute Human APEG1 to 2 μg/mL in 1X PBST, add 100 μL/well, and incubate overnight at 2-8°C.
2. Washing: Wash the plate with wash buffer (260 μL/well) , once, and pat dry.
3. Blocking: Add 150 μL/well of blocking buffer, incubate at 25°C with shaking at 450 rpm for 4 hours.
4. Dilute the primary antibody to concentrations of 0, 0.005, 0.01, 0.05, 0.1, 0.5, 1, 5, 10, 20, 50, 100, and 250 μg/mL.
5. Add primary antibody: 100 μL/well. Incubate for 2 hours at 25°C, 450 rpm.
6. Wash plates: Wash plates with wash buffer, 260 μL/well. Wash five times, allowing each wash to stand for 1 minute, then blot dry.
7. Add secondary antibody: Dilute secondary antibody at a 1:2000 ratio. Add 100 μL per well and incubate at 25°C, 450 rpm for 1 hour.
8. Washing: Wash the plate with wash buffer (260 μL/well) , three times, with each wash lasting 1 minute, then pat dry.
9. Add TMB Substrate Solution: Add 100 μL/well, incubate at room temperature in the dark for 15 minutes.
10. Stop Reaction: Add 100 μL of 2 M H₂SO₄ per well.
11. Read Plate: Measure absorbance at 450 nm.
12. Using GraphPad Prism software, plot a curve with the OD value as the vertical axis and the Anti-APEG1 Antibody concentration as the horizontal axis, and calculate the ED₅₀ value.

Human

E. coli

C-6*His

Q15772-4 (M1-E113)

10290  [NCBI]

Full Length of Isoform-4

MKPSPSQNRRSSDTGSKAPPTFKVSLMDQSVREGQDVIMSIRVQGEPKPVVSWLRNRQPVRPDQRRFAEEAEGGLCRLRILAAERGDAGFYTCKAVNEYGARQCEARLEVRGE

Approximately 15 kDa, based on SDS-PAGE under reducing conditions.

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  ≥ 95%, as determined by reducing SDS-PAGE.

Solution.

Supplied as a 0.22 μm filtered solution of PBS, pH 7.4.

<1 EU/μg, determined by LAL method.

N/A.

Stored at -80°C for 1 year from date of receipt. It is stable at -20°C for 3 months after opening. It is recommended to freeze aliquots at -80°C for extended storage. Avoid repeated freeze-thaw cycles.

Shipping with dry ice.