In-Cell ELISAs

In-Cell ELISA, also known as cell-based ELISA, In-Cell western or cytoblot, delivers quantitative data of intracellular signaling proteins without the need to lyse cells or run gels. 

​​​​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.

  • ​​​​​Cells are analyzed in situ eliminating sample preparation steps like cell lysis.


Quantitative – antibodies are labeled to enable detection using a microplate reader (absorbance or fluorescence).

  • Cells can be counter-stained with Janus Green which allows normalization to cell number.

Broad dynamic range – accurate quantification without the need for many wells with different cell densities.

  • Figure 1 shows that differences in enzyme levels in a particular cell line can be more accurately detected using In-Cell ELISA, compared to western blot. 

ab110289 in cell elisa v2

​​​​​​​​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 shows mass spectrometry data used to confirm the binding specificity of antibodies characterized for use in In-Cell ELISA kits. 
ab110292 in cell elisa v2 mass spectrometry

​​​​​​​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 nameProduct nameDetection methodFluorescence (Ex/Em)Product 
AKT (Total and pS473)AKT Total + pS473 In-Cell ELISA Colorimetric
AKT (Total and pS473)AKT Total + pS473 In-Cell ELISA Kit IR
AKT (Total and pS473)AKT Total + pSer473 In-Cell ELISA Kit 


AP: 360/449 nm HRP: 555/595 nm

Cleaved PARPCleaved PARP Human In-Cell ELISA Kit Colorimetric
Cleaved PARPCleaved PARP Human In-Cell ELISA Kit FluorescentHRP: 555/595 nmab140362
Cleaved PARPCleaved PARP Human In-Cell ELISA KitIR

EGFR (Tyr1068)
EGFR (Tyr1068) Human In-Cell ELISA Kit Colorimetric
EGFR (Tyr845)

EGFR (Tyr845) In-Cell ELISA Kit

EGFR (Tyr992)EGFR (Tyr992) In-Cell ELISA Kit Colorimetric

EGFR Human In-Cell ELISA Kit

ERK1/2 (Thr202/Tyr204)

ERK1/2 (Thr202/Tyr204) In-Cell ELISA Kit

Fatty AcidFatty Acid Oxidation Human In-Cell ELISA Kit  Colorimetric
JNK (Thr183/Tyr185)JNK (Thr183/Tyr185) In-Cell ELISA Kit Colorimetric
p38 MAPK alpha (Thr180/Tyr182)p38 MAPK alpha (Thr180/Tyr182) In-Cell ELISA Kit Colorimetric
PhosphotyrosinepTyr In-Cell ELISA Kit Colorimetric
RSK1 p90 (Total and pSer380)RSK1 p90 (Total and pSer380) In-Cell ELISA KitFluorescentAP: 360/449 nm HRP: 555/595 nmab170196
SIRT1 (Total and pSer47)SIRT1 (Total and pSer47) Human In-Cell ELISA Kit Colorimetric
SIRT1 (Total and pSer47)SIRT1 (Total and pSer47) Human In-Cell ELISA Kit FluorescentAP: 360/449 nm HRP: 555/595 nmab136808
STAT1 (Tyr701)STAT1 (Tyr701) In-Cell ELISA Kit Colorimetric
STAT3 (Tyr705)

STAT3 (Tyr705) In-Cell ELISA Kit

STAT4 (Tyr693)

STAT4 (Tyr693) In-Cell ELISA Kit

STAT5 (Tyr694)

STAT5 (Tyr694) In-Cell ELISA Kit

STAT6 (Tyr641)

STAT6 (Tyr641) In-Cell ELISA Kit

-Cell Cycle In-Cell ELISA KitFluorescentAP: 360/449 nm HRP: 555/595 nmab140363
-MitoBiogenesis™ In-Cell ELISA Kit FluorescentAP: 360/449 nm HRP: 555/595 nmab140359
-MitoBiogenesis™ In-Cell ELISA Kit Colorimetric

​Find a dual target detection kit with IR readout below:

Target nameProduct nameProduct code
AMPK alpha 1 (Total and pT172)AMPKa (Total and pT172) In-Cell ELISA Kit ab151280
H2A.X (pSer139) and Cleaved-PARPH2A.X (pSer139) + Cleaved-PARP Human In-Cell ELISA Kitab131383
Hif1 and ActinHif1 + Actin Hypoxia Human In-Cell ELISA Kit ab125300
Hif1 and PDK1Hif1 + PDK1 Hypoxia Human In-Cell ELISA Kitab125299
Hif1 alpha and GLUTHif1 alpha + GLUT Hypoxia Human In-Cell ELISA Kit ab125298
Hif1 alpha and BNIP3Hif1 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
PhosphoPDHpPDH 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 Kitab135394
-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.