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This assay works like an ELISA by using an antibody specific to your target of interest in fixed, permeabilized cells. Single targets can be detected by colorimetric, fluorescent or infra-red (IR) readouts; dual targets can be detected by IR fluorescence.
In-Cell ELISA offers several distinct advantages over western blot:
Simple – minimal hands-on time.
Quantitative – antibodies are labeled to enable detection using a microplate reader (absorbance or fluorescence).
Broad dynamic range – accurate quantification without the need for many wells with different cell densities.
Figure 1: In-Cell ELISA detects higher levels of ACAA1 compared to western blot. Human fibroblast cell lines deficient in various enzymes involved fatty acid beta-oxidation were screened in western blot and In-Cell ELISA. Western blot signal looked very similar in all cell line in the right section of the blot. However, In-Cell ELISA data showed that cell line 482 that had higher levels of ACAA1 (using characterized antibody to ACAA1 (ab110289)). (Figure 1A - molecular weight markers (top to bottom): 250, 150, 100, 75, 50, 37, 25, 20, 15,10 kDa).
Specific - antibodies used in In-Cell ELISA and those recommended for use with In-Cell ELISA support pacts are rigorously tested for specificity. The antibodies are characterized using immunoprecipitation, mass spectrometry, western blotting and fluorescence immunocytochemistry.
Figure 2: Immunoprecipitation and mass spectrometry confirm the specifity of antibody to catalase (ab110292). During development of In-Cell ELISA catalase antibody ab110292, multiple hybridoma cells lines were used to precipitate catalase from liver tissue extract. A single band at 60 kDa was identified (the band at 150 kDa is IgG). The 60 kDa band was cut out, trypsinized and analyzed by tandem mass spectrometry, and was found to be consistent with human catalase.
Flexible - In-Cell ELISA kits can be used with cultured adherent or suspension cells. Cells are fixed, permeabilized and the intracellular targets of interest are detected by antibodies, which are subsequently detected with labeled secondary antibodies.
Can't find a kit to your protein of interest? Use your own antibodies with our In-Cell ELISA Support Packs.
See all of our In-Cell ELISA kits here
|Target name||Product name||Detection method||Fluorescence (Ex/Em)||Product|
|AKT (Total and pS473)||AKT Total + pS473 In-Cell ELISA||Colorimetric||ab126578|
|AKT (Total and pS473)||AKT Total + pS473 In-Cell ELISA Kit||IR||ab126579|
|AKT (Total and pS473)||AKT Total + pSer473 In-Cell ELISA Kit|
AP: 360/449 nm HRP: 555/595 nm
|Cleaved PARP||Cleaved PARP Human In-Cell ELISA Kit||Colorimetric||ab139410|
|Cleaved PARP||Cleaved PARP Human In-Cell ELISA Kit||Fluorescent||HRP: 555/595 nm||ab140362|
|Cleaved PARP||Cleaved PARP Human In-Cell ELISA Kit||IR||ab110215|
|EGFR (Tyr1068) Human In-Cell ELISA Kit||Colorimetric||ab126420|
|EGFR (Tyr992)||EGFR (Tyr992) In-Cell ELISA Kit||Colorimetric||ab126422|
|Fatty Acid||Fatty Acid Oxidation Human In-Cell ELISA Kit||Colorimetric||ab118182|
|JNK (Thr183/Tyr185)||JNK (Thr183/Tyr185) In-Cell ELISA Kit||Colorimetric||ab126424|
|p38 MAPK alpha (Thr180/Tyr182)||p38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit||Colorimetric||ab126425|
|Phosphotyrosine||pTyr In-Cell ELISA Kit||Colorimetric||ab126431|
|RSK1 p90 (Total and pSer380)||RSK1 p90 (Total and pSer380) In-Cell ELISA Kit||Fluorescent||AP: 360/449 nm HRP: 555/595 nm||ab170196|
|SIRT1 (Total and pSer47)||SIRT1 (Total and pSer47) Human In-Cell ELISA Kit||Colorimetric||ab136807|
|SIRT1 (Total and pSer47)||SIRT1 (Total and pSer47) Human In-Cell ELISA Kit||Fluorescent||AP: 360/449 nm HRP: 555/595 nm||ab136808|
|STAT1 (Tyr701)||STAT1 (Tyr701) In-Cell ELISA Kit||Colorimetric||ab126426|
|-||Cell Cycle In-Cell ELISA Kit||Fluorescent||AP: 360/449 nm HRP: 555/595 nm||ab140363|
|-||MitoBiogenesis™ In-Cell ELISA Kit||Fluorescent||AP: 360/449 nm HRP: 555/595 nm||ab140359|
|-||MitoBiogenesis™ In-Cell ELISA Kit||Colorimetric||ab110217|
Find a dual target detection kit with IR readout below:
|Target name||Product name||Product code|
|AMPK alpha 1 (Total and pT172)||AMPKa (Total and pT172) In-Cell ELISA Kit||ab151280|
|H2A.X (pSer139) and Cleaved-PARP||H2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kit||ab131383|
|Hif1 and Actin||Hif1 + Actin Hypoxia Human In-Cell ELISA Kit||ab125300|
|Hif1 and PDK1||Hif1 + PDK1 Hypoxia Human In-Cell ELISA Kit||ab125299|
|Hif1 alpha and GLUT||Hif1 alpha + GLUT Hypoxia Human In-Cell ELISA Kit||ab125298|
|Hif1 alpha and BNIP3||Hif1 alpha + BNIP3 Hypoxia Human In-Cell ELISA Kit||ab129733|
|mTOR (Total and pSer2448)||mTOR (pSer2448) In-Cell ELISA Kit||ab168539|
|p53 (Total and pSer392)||p53 Total + pSer392 Human In-Cell ELISA Kit||ab128571|
|p53 (Total and pSer46)||p53 Total + pSer46 Human In-Cell ELISA Kit||ab128570|
|p53 (Total and pSer9)||p53 Total + pSer9 Human In-Cell ELISA Kit||ab135393|
|PhosphoPDH||pPDH In-Cell ELISA Kit||ab110218|
|RSK1 p90 (Total and pSer380)||RSK1 p90 (Total and pSer380) Human In-Cell ELISA Kit||ab157398|
|SIRT1 (Total and pSer47)||SIRT1 Total and pSer47 Human In-Cell ELISA Kit||ab135394|
|-||Cell Cycle Human In-Cell ELISA Kit||ab139412|
|-||MitoBiogenesis™ In-Cell ELISA Kit||ab110216|
If you have antibodies to your target that you would like to use, In-Cell ELISA Support Packs are available which contain all the pre-made buffers for performing the assay with adherent and non-adherent cells. When using cells that may detach due to the specific treatment you are using e.g., apoptotic, detaching cells, use our specific In-Cell ELISA Support Pack (ab111542) which includes assay plates, reagents and a protocol.