ERK1, JNK, P38 ELISA Kit

Catalog No. ABIN4886189

The Human, Mouse and Rat ERK1, JNK, P38 ELISA Kit (ABIN4886189) is a Colorimetric ELISA Kit designed to quantify Human, Mouse and Rat ERK1, JNK, P38. This ELISA Kit has been cited in 2 publications.

Quick Overview for ERK1, JNK, P38 ELISA Kit (ABIN4886189)

Target: ERK1, JNK, P38

Binding Specificity: pThr180, pThr183, pThr202, pTyr182, pTyr185, pTyr204

Reactivity: Human, Mouse, Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Application: ELISA

Sample Type: Cell Lysate, Tissue Lysate

Product Details

Purpose: Phospho-ERK/JNK/P38? ELISA Kit. This assay semi-quantitatively measures phophorylated ERK/JNK/P38? in lysate samples.

Analytical Method: Semi-Quantitative

Specificity: The antibody pair provided in this kit recognizes ERK1/2 (T202/Y204), JNK (T183/Y185), P38α (T180/Y182), Total ERK1/2, JNK and P38α in Human, Mouse and Rat Cell Lysates.

Characteristics:

• Simultaneously measure Phosphorylated protein and pan protein in one experiment (for normalization purpose)
• Screen numerous different cell lysates without performing a Western Blot analysis
• Minimal hands-on time, convenient, and non-radioactive material

Components:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Anti-Phospho Antibody
• HRP-Conjugated Secondary Antibody
• Assay Diluent
• TMB One-Step Substrate
• Stop Solution
• Lysis Buffer
• Positive Control Sample

Material not included:

• Distilled or deionized water
• 100 mL and 1 liter graduated cylinders
• Tubes to prepare sample dilutions
• Protease and Phosphatase inhibitors
• Precision pipettes to deliver 2 μL to 1 mL volumes
• Adjustable 1-25 mL pipettes for reagent preparation
• Benchtop rocker or shaker
• Microplate reader capable of measuring absorbance at 450 nm

Application Details

Sample Volume: 100 μL

Plate: Pre-coated

Protocol:

1. Prepare all reagents and samples as instructed in the manual.
2. Add 100 μL of sample or positive control to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared primary antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared 1X HRP-Streptavidin to each well.
7. Incubate 1 h at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Restrictions: For Research Use only

Handling

Storage: -20 °C

Storage Comment: Upon receipt, the kit should be stored at -20 °C. Please use within 6 months from the date of shipment. After initial use, Wash Buffer Concentrate (Item B), Assay Diluent (Item E), TMB One-Step Substrate Reagent (Item H), HRP-Streptavidin (Item G), Stop Solution (Item I) and Cell Lysate Buffer (Item J) should be stored at 4 °C to avoid repeated freeze-thaw cycles. Return unused wells to the pouch containing desiccant pack, reseal along entire edge and store at -20 °C. Reconstituted Positive Control (Item K) should be stored at -70 °C.

Expiry Date: 6 months

References

Elamin, Masclee, Troost, Pieters, Keszthelyi, Aleksa, Dekker, Jonkers: "Ethanol impairs intestinal barrier function in humans through mitogen activated protein kinase signaling: a combined in vivo and in vitro approach." in: PLoS ONE, Vol. 9, Issue 9, pp. e107421, (2015) (PubMed).

Elsherbiny, Abd El Galil, Gabr, Al-Gayyar, Eissa, El-Shishtawy: "Reno-protective effect of NECA in diabetic nephropathy: implication of IL-18 and ICAM-1." in: European cytokine network, Vol. 23, Issue 3, pp. 78-86, (2012) (PubMed).

Target Details

Target: ERK1, JNK, P38

Alternative Name: ERK1, JNK , P38