Product Pathways - PathScan ELISA
PathScan® Total α-Tubulin Sandwich ELISA Kit #7944
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|Kit Includes||Volume||Solution Color|
|Alpha-Tubulin Rabbit Antibody Coated Microwells||96 tests|
|Alpha-Tubulin Mouse Detection Ab||11 ml||Green|
|Anti-mouse 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 R Hm Mk
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
The PathScan® Total α-Tubulin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of α-tubulin. An α-tubulin rabbit antibody has been coated onto the microwells. After incubation with cell lysates, α-tubulin is captured by the coated antibody. Following extensive washing, an α-tubulin mouse detection antibody is added to detect the captured α-tubulin. An anti-mouse IgG, HRP-linked antibody 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 α-tubulin.Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Total α-Tubulin Sandwich ELISA Kit detects endogenous levels of α-tubulin protein, as shown in Figure 1. The kit sensitivity is shown in Figure 2. Microtubule stabilizing or destabilizing agents may significantly increase or decrease the signal, respectively. 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 A-431 cells with EGF stimulates phosphorylation of the EGF receptor at Tyr1086 as detected by PathScan® Phospho-EGF Receptor (Tyr1086) Sandwich ELISA Kit #7240, but does not affect the level of α-tubulin as detected by PathScan® Total α-Tubulin Sandwich ELISA Kit #7944. A-431 cells were treated with EGF (100 ng/ml, 3 min.) at 37ºC before lysis. Absorbance at 450 nm is shown in the top figure, while the corresponding western blots using Total α-Tubulin (11H10) Rabbit mAb #2125 (left) or Phospho-EGF Receptor (Tyr1086) Antibody #2220 (right) 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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- 7945 PathScan® Total α-Tubulin Sandwich ELISA Antibody Pair
- 2125 α-Tubulin (11H10) Rabbit mAb
- 2144 α-Tubulin Antibody
- 3873 α-Tubulin (DM1A) Mouse mAb
- 2148 α/β-Tubulin Antibody
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 9803 Cell Lysis Buffer (10X)
- 7004 TMB Substrate
- 7002 STOP Solution
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
For Research Use Only. Not For Use In Diagnostic Procedures.