Product Pathways - PathScan ELISA
PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit #7204
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|Kit Includes||Volume||Solution Color|
|Tubulin Ab-Coated Microwells||96 tests|
|Acetylated-Lysine Detection Ab||11 ml||Green|
|Anti-rabbit IgG, HRP-linked Antibody||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules –Each module is designed to break apart for 8 tests.
Note: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
H M Mk
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
CST's PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of acetylated α-tubulin protein. An α-tubulin mouse mAb has been coated onto the microwells. After incubation with cell lysates, α-tubulin protein (acetylated and non-acetylated) is captured by the coated antibody. Following extensive washing, an acetyl-lysine rabbit Ab is added to detect the acetylated α-tubulin protein. Anti-rabbit IgG, HRP-linked Antibody #7074 is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of acetylated α-tubulin protein.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Acetyl-α-Tubulin Sandwich ELISA Kit #7204 detects endogenous levels of acetylated α-tubulin. As shown in Figure 1 using the Acetyl-α-Tubulin Sandwich ELISA Kit #7204, a high level of acetylated α-tubulin is detected in COS cells when treated with TSA. The level of total α-tubulin (acetylated and non-acetylated) remains unchanged as shown by Western analysis (Figure 1). Similar results are obtained when NIH/3T3 and Jurkat cells are treated with TSA (data not shown). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1. Treatment of COS cells with Trichostatin A (TSA) increases the acetylation of α-tubulin detected by PathScan® Acetyl-alpha-Tubulin Sandwich ELISA Kit #7204. TSA treatment does not affect the level of α-tubulin that is detected by Western analysis. COS cells (70-80% confluent) were treated for 16-18 hours with 0.4 micromolar TSA at 37oC. Absorbance readings at 450 nm are shown in the top figure while the corresponding Western blots using α-Tubulin Antibody #2125 (left panel) or Acetylated-Lysine Antibody #9441 (right panel) are shown in the bottom figure.
The cytoskeleton consists of three types of cytosolic fibers: microtubules, microfilaments (actin filaments), and intermediate filaments. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. γ-tubulin is required to nucleate polymerization of tubulin subunits to form microtubule polymers. Many cell movements are mediated by microtubule action, including the beating of cilia and flagella, cytoplasmic transport of membrane vesicles, chromosome alignment during meiosis/mitosis, and nerve-cell axon migration. These movements result from competitive microtubule polymerization and depolymerization or through the actions of microtubule motor proteins (1).
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- 7944 PathScan® Total α-Tubulin Sandwich ELISA Kit
- 2144 α-Tubulin Antibody
- 2125 α-Tubulin (11H10) Rabbit mAb
- 9441 Acetylated-Lysine Antibody
- 9681 Acetylated-Lysine Mouse mAb (Ac-K-103)
- 9803 Cell Lysis Buffer (10X)
- 9950 Trichostatin A (TSA)
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.