Anti-ICAM1 antibody [EPR22161-284] (ab222736)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR22161-284] to ICAM1
- Suitable for: WB, IHC-P, Flow Cyt, ICC/IF, IP
- Reacts with: Mouse
Overview
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Product name
Anti-ICAM1 antibody [EPR22161-284]
See all ICAM1 primary antibodies -
Description
Rabbit monoclonal [EPR22161-284] to ICAM1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species Flow Cyt MouseICC/IF MouseIHC-P MouseIP MouseWB Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse kidney, spleen and liver tissue lysate; A20 whole cell lysate. IHC-P: Mouse lung and spleen tissue. ICC/IF: A20 cells. Flow: Mouse primary splenocytes. IP: Mouse spleen lysate.
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General notes
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 production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR22161-284 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-ICAM1 antibody [EPR22161-284] (ab222736) at 1/1000 dilution
Lane 1 : Mouse kidney tissue lysate
Lane 2 : Mouse spleen tissue lysate
Lane 3 : Mouse liver tissue lysate
Lane 4 : A20 (mouse reticulum sarcoma b lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure time: 48 seconds.
The 110-KDa apparent molecular mass is due to glycosylation (PMID: 1680698).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A20 (mouse reticulum sarcoma cell line) cells labeling ICAM with ab222736 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in A20 cells with distinct staining of the target protein being transported from Golgi to cell surfaces (PMID: 17486117) is observed. Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used as a countertsain (Red) at 1/200 dilution. The nuclear counterstain is DAPI (Blue).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Alexa Fluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution.
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling ICAM1 using ab222736 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on the marginal zone of mouse spleen (PMID: 12130787; PMID: 17947659) is observed. Counterstained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunostaining was performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab222736 for 30 mins at room temperature.
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ICAM1 was immunoprecipitated from 0.35 mg Mouse spleen lysate with ab222736 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222736 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/5000 dilution.
Lane 1: Mouse spleen tissue lysate 10 μg (input).
Lane 2: ab222736 IP in mouse spleen tissue lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222736 in Mouse spleen lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.110-kDa molecular mass is due to glycosylation (PMID: 1680698).
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Flow cytometric analysis of Mouse primary splenocytes labeling ICAM1 with ab222736 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabeled control (Cell without incubation with primary antibody and secondary antibody, Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/200 dilution was used as the secondary antibody. Gated on viable cells.
ICAM1 negative population (left half of red peak) was also observed in mouse splenocytes which is consistent with IHC staining pattern.
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Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling ICAM1 using ab222736 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous and cytoplasmic staining on mouse lung (PMID: 14568945; PMID: 9802985) is observed. Counterstained with DAPI.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) ready to use.
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
Immunostaining was performed on a Leica Biosystems BOND® RX instrument. The section was incubated with ab222736 for 30 mins at room temperature.
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