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Rabbit Recombinant Monoclonal ERK1 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Synthetic peptide - Human samples. Cited in 72 publications.


Images

Immunoprecipitation - Anti-ERK1 antibody [Y72] (AB32537), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (AB32537), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (AB32537), expandable thumbnail
  • Western blot - Anti-ERK1 antibody [Y72] (AB32537), expandable thumbnail
  • Western blot - Anti-ERK1 antibody [Y72] (AB32537), expandable thumbnail

Publications

Key facts

Isotype
IgG
Host species
Rabbit
Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form
Liquid
Clonality
Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Expected
Expected
Tested
Expected
Expected
Synthetic peptide - Human
Not recommended
Not recommended
Tested
Not recommended
Not recommended

Tested
Tested

Species
Human
Dilution info
2 µg/mL
Notes

Perform heat-mediated antigen retrieval before commencing with IHC staining protocol.

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/20 - 1/40
Notes

-

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Synthetic peptide - Human
Dilution info
1/1000
Notes

-

Species
Mouse
Dilution info
1/1000
Notes

Can be blocked with ab204281.

Species
Human
Dilution info
1/1000
Notes

-

Tested
Tested

Species
Human
Dilution info
1/400
Notes

This product gave a positive signal in HAP1 cells fixed with 4% formaldehyde (10 min) and 100% methanol (5 min).

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Tested
Tested

Species
Human
Dilution info
1/30 - 1/40
Notes

Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Expected
Expected

Species
Mouse
Dilution info
Use at an assay dependent concentration.
Notes

-

Not recommended
Not recommended

Species
Synthetic peptide - Human
Dilution info
-
Notes

-

Associated Products

Select an associated product type

10 products for Alternative Product

Target data

Function

Serine/threonine kinase which acts as an essential component of the MAP kinase signal transduction pathway (PubMed:34497368). MAPK1/ERK2 and MAPK3/ERK1 are the 2 MAPKs which play an important role in the MAPK/ERK cascade. They participate also in a signaling cascade initiated by activated KIT and KITLG/SCF. Depending on the cellular context, the MAPK/ERK cascade mediates diverse biological functions such as cell growth, adhesion, survival and differentiation through the regulation of transcription, translation, cytoskeletal rearrangements. The MAPK/ERK cascade also plays a role in initiation and regulation of meiosis, mitosis, and postmitotic functions in differentiated cells by phosphorylating a number of transcription factors. About 160 substrates have already been discovered for ERKs. Many of these substrates are localized in the nucleus, and seem to participate in the regulation of transcription upon stimulation. However, other substrates are found in the cytosol as well as in other cellular organelles, and those are responsible for processes such as translation, mitosis and apoptosis. Moreover, the MAPK/ERK cascade is also involved in the regulation of the endosomal dynamics, including lysosome processing and endosome cycling through the perinuclear recycling compartment (PNRC); as well as in the fragmentation of the Golgi apparatus during mitosis. The substrates include transcription factors (such as ATF2, BCL6, ELK1, ERF, FOS, HSF4 or SPZ1), cytoskeletal elements (such as CANX, CTTN, GJA1, MAP2, MAPT, PXN, SORBS3 or STMN1), regulators of apoptosis (such as BAD, BTG2, CASP9, DAPK1, IER3, MCL1 or PPARG), regulators of translation (such as EIF4EBP1) and a variety of other signaling-related molecules (like ARHGEF2, DEPTOR, FRS2 or GRB10) (PubMed:35216969). Protein kinases (such as RAF1, RPS6KA1/RSK1, RPS6KA3/RSK2, RPS6KA2/RSK3, RPS6KA6/RSK4, SYK, MKNK1/MNK1, MKNK2/MNK2, RPS6KA5/MSK1, RPS6KA4/MSK2, MAPKAPK3 or MAPKAPK5) and phosphatases (such as DUSP1, DUSP4, DUSP6 or DUSP16) are other substrates which enable the propagation the MAPK/ERK signal to additional cytosolic and nuclear targets, thereby extending the specificity of the cascade.

Alternative names

Recommended products

Rabbit Recombinant Monoclonal ERK1 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Synthetic peptide - Human samples. Cited in 72 publications.

Key facts

Isotype
IgG
Form
Liquid
Clonality
Monoclonal
Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number
Y72
Purification technique
Affinity purification Protein A
Specificity

This antibody recognises ERK1.The antibody does not cross-react with other MAP kinases.

Epitope
ab32537 reacts with an epitope located in the N terminal region of ERK1.
Concentration
Loading...

Storage

Shipped at conditions
Blue Ice
Appropriate short-term storage duration
1-2 weeks
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
-20°C
Aliquoting information
Upon delivery aliquot
Storage information
Avoid freeze / thaw cycle

Notes

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.
Activity summary

ERK1 also known as MAPK3 is an extracellular signal-regulated kinase involved in transmitting signals from the cell surface to the nucleus. This protein has a molecular mass of about 44 kDa. ERK1 expresses in various tissue types with higher expression in the brain heart and skeletal muscle. Researchers often study ERK1 in the context of its role in cellular signaling due to its involvement in critical regulatory functions.

Biological function summary

ERK1 plays a significant role in cell cycle regulation differentiation and proliferation. It forms part of the MAPK signaling cascade becoming activated through a phosphorylation event. In its activated form ERK1 translocates to the nucleus where it phosphorylates target substrates. ERK1 often functions in conjunction with its homolog ERK2 to mediate these cellular processes marking it as an essential player in growth factor signaling.

Pathways

ERK1 functions primarily within the MAPK/ERK signaling pathway a major conduit for transmitting proliferative signals from growth factor receptors. ERK1 interacts with proteins like MEK1/2 which phosphorylate and activate ERK1 in response to extracellular stimuli. Another critical pathway involving ERK1 is the Ras-Raf-MEK-ERK cascade which regulates various cellular outcomes. This connection to the Ras family highlights its importance in signal transduction and reinforces its position in critical cellular processes.

Associated diseases and disorders

Aberrant activation of ERK1 connects to diseases such as cancer and cardiovascular disorders. In cancer the dysregulation of the MAPK/ERK pathway often through mutations affecting Ras or Raf proteins leads to uncontrolled cell proliferation. ERK1's involvement in cardiovascular diseases links to its role in hypertrophic signaling in cardiac cells where altered ERK1 activity can contribute to pathological cardiac remodeling. Understanding these interactions can aid in developing therapeutic strategies targeting the MAPK/ERK signaling pathway.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

19 product images

  • Immunoprecipitation - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Immunoprecipitation - Anti-ERK1 antibody [Y72] (ab32537)

    ab32537 (purified) at a dilution of 1/20 immunoprecipitating ERK1 in Jurkat whole cell lysate.

    Lane 1 (input): Jurkat whole cell lysate (10μg)

    Lane 2 (+): ab32537 + Jurkat whole cell lysate.

    Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32537 in Jurkat whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) ab131366 VeriBlot for IP (HRP) was used for detection (1/10000).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Immunoprecipitation - Anti-ERK1 antibody [Y72] (ab32537)

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (ab32537)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cervix carcinoma tissue labelling ERK1 with purified ab32537 at a dilution of 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (ab32537)

    ab32537 staining ERK1 in wild-type HAP1 cells (top panel) and ERK1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab32537 at 1/400 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1ug/ml concentration overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed ab150117) at 2ug/ml (shown in pseudo-color red). Nuclear DNA was labelled in blue with DAPI.

    This product also gave a positive signal under the same testing conditions in HAP1 cells fixed with 4% formaldehyde (10 min).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 μg)
    Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 μg)
    Lanes 1 and 2: Green signal from target - ab32537 observed at 42 kDa
    Lanes 3 and 4: Red signal from loading control - Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 observed at 42 kDa
    Lanes 5 and 6: Merged (red and green) signal
    ab32537 was shown to specifically react with ERK1 when ERK1 knockout samples were used.
    Wild-type and ERK1 knockout samples were subjected to SDS-PAGE. ab32537 and Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 (loading control to beta actin) were both diluted 1/1000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) andGoat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Predicted band size: 43 kDa

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Lanes 1- 2: Merged signal (red and green). Green - ab32537 observed at 43 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.

    ab32537 was shown to react with ERK1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human MAPK3 (ERK1) knockout HEK-293T cell line ab266519 (knockout cell lysate Human MAPK3 (ERK1) knockout HEK-293T cell lysate ab257099) was used. Wild-type HEK-293T and MAPK3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32537 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: MAPK3 knockout HEK-293T cell lysate at 20 µg

    Lane 2: Western blot - Human MAPK3 (ERK1) knockout HEK-293T cell line (Human MAPK3 (ERK1) knockout HEK-293T cell line ab266519)

    Performed under reducing conditions.

    Predicted band size: 43 kDa

    Observed band size: 43 kDa

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Lanes 1, 3 and 5: Wild-type HAP1 cell lysate (20 μg)
    Lanes 2, 4 and 6: ERK1 knockout HAP1 cell lysate (20 μg)
    Lanes 1 and 2: Green signal from target
    Lanes 3 and 4: Red signal from loading control
    Lanes 5 and 6: Merged (red and green) signal

    Red - loading control, Anti-beta Actin antibody [mAbcam 8226] - Loading Control ab8226 observed at 42 kDa or Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa

    This western blot image is a comparison between ab32537 and a competitor's top cited rabbit polyclonal antibody.

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Predicted band size: 43 kDa

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Blocking and dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537) at 1/10000 dilution

    Lane 1: 293T whole cell lysate at 10 µg

    Lane 2: HeLa whole cell lysate at 10 µg

    Lane 3: Jurkat whole cell lysate at 10 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

  • Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-MAPK3 knockout cells (red line) stained with ab32537. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab32537, 1μg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. A Rabbit IgG isotype control antibody (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-MAPK3 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Blocking and dilution buffer: 5% NFDM/TBST

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution

    All lanes: RAW264.7 whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/10000 dilution

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

  • Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537)

    Intracellular Flow Cytometry analysis ofJurkat cells labelling ERK1 with purified ab32537 at a dilution of 1/30 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab32537 overnight at 4°C. Antibody binding was detected using a goat anti-rabbit Alexa Fluor® 790 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution

    Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

    Lane 2: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

    Lane 3: Recombinant Human ERK1 protein (ab43623) (ab43623) at 20 µg

    Lane 4: Western blot - Recombinant Human ERK2 protein (Recombinant Human ERK2 protein ab43625) at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) ab175781) at 1/10000 dilution

    Predicted band size: 43 kDa

    Observed band size: 44 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (ab32537)

    Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling ERK1 with purified ab32537 at a dilution of 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution

    All lanes: Jurkat cell lysate

    Predicted band size: 43 kDa

    Observed band size: 43 kDa

  • Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537)

    Overlay histogram showing HeLa cells stained with ab32537 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween 20 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32537, 1/11312) for 30 min at 22°C. The secondary antibody used was Alexa Fluorr® 488 goat anti-rabbit IgG (H&L) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, 0.01μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.

    This antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween 20 for 20 min used under the same conditions.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (ab32537)

    IHC image of ab32537 staining ERK1 in Human tonsil formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32537, 2μg/ml dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail
    This image is courtesy of a customer review submitted by Kirk McManus.

    Immunocytochemistry/ Immunofluorescence - Anti-ERK1 antibody [Y72] (ab32537)

    Unpurified ab32537 staining ERK1 in HeLa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, a goat anti rabbit Alexa Fluor® 488 (1/200) was used as the secondary antibody. Counterstained with DAPI. Cytoplasmic and nuclear staining shown.

  • Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail
    This image is courtesy of an anonymous customer review

    Flow Cytometry (Intracellular) - Anti-ERK1 antibody [Y72] (ab32537)

    Unpurified ab32537 staining ERK1 (green) in HEK293 cells by intracellular flow cytometry. Cells were fixed with paraformaldehyde and permeabilized with 70% methanol. The sample was incubated with the primary antibody (1/40 in PBS + 0.2% BSA + 0.1% sodium azide) for 1 hour at 22°C. A phycoerythrin-conjugated goat anti-rabbit IgG (1/100) was used as the secondary antibody. Gating Strategy: Live Cells. Purple plot represents isotype control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ERK1 antibody [Y72] (ab32537)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue labelling ERK1 with unpurified ab32537.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Western blot - Anti-ERK1 antibody [Y72] (ab32537), expandable thumbnail

    Western blot - Anti-ERK1 antibody [Y72] (ab32537)

    Blocking buffer and concentration: 5% NFDM/TBST. 

    Diluting buffer and concentration: 5% NFDM/TBST.

    This blot was developed using a high sensitivity ECL substrate.

    Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a loading control.

    All lanes: Western blot - Anti-ERK1 antibody [Y72] (ab32537) at 1/1000 dilution

    Lane 1: Mouse brain lysate at 20 µg

    Lane 2: Mouse heart lysate at 20 µg

    Lane 3: Mouse kidney lysate at 20 µg

    Lane 4: Mouse spleen lysate at 20 µg

    Lane 5: Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

    Lane 6: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 43 kDa

    Observed band size: 43 kDa

    Exposure time: 180s

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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