Overview

  • Product name
    Anti-ICAM1 antibody [MEM-111]
    See all ICAM1 primary antibodies
  • Description
    Mouse monoclonal [MEM-111] to ICAM1
  • Host species
    Mouse
  • Specificity
    This antibody reacts with human CD54 antigen.
  • Tested applications
    Suitable for: ICC/IF, ELISA, WB, IHC-P, IHC-Fr, Flow Cyt, Sandwich ELISAmore details
  • Species reactivity
    Reacts with: Cow, Human
  • Immunogen

    Tissue/ cell preparation (Human). (Burkitt's lymphoma cell line RAJI).

  • Positive control
    • For WB it is highly recommended to use TNF-alpha activated HUVEC cells as a positive control. Sandwich ELISA: supernatant of HUVEC cell line. Indirect ELISA: endothelial cell line derived from bovine pulmonary artery. RAJI cell line,K562 leukemia cell line, JY cell line, Activated T-lymphocytes, Thymus, RE cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab2213 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/100.
ELISA Use a concentration of 25 µg/ml. Use at a dilution of 1/1000 for Indirect ELISA, 1hr, RT. The antibody reacts with tissue - endothelial cell line derived from bovine pulmonary artery.
WB Use at an assay dependent concentration. Use under non-reducing conditions.
IHC-P Use a concentration of 10 µg/ml.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use a concentration of 2 µg/ml.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

Sandwich ELISA Use a concentration of 5 µg/ml. Can be paired for Sandwich ELISA with Rabbit polyclonal to ICAM1 (Biotin) (ab7815).

For sandwich ELISA, use this antibody as Capture at 5µg/ml with ab7815 as Detection.

Target

  • Function
    ICAM proteins are ligands for the leukocyte adhesion protein LFA-1 (integrin alpha-L/beta-2). During leukocyte trans-endothelial migration, ICAM1 engagement promotes the assembly of endothelial apical cups through ARHGEF26/SGEF and RHOG activation. In case of rhinovirus infection acts as a cellular receptor for the virus.
  • Sequence similarities
    Belongs to the immunoglobulin superfamily. ICAM family.
    Contains 5 Ig-like C2-type (immunoglobulin-like) domains.
  • Post-translational
    modifications
    Monoubiquitinated, which is promoted by MARCH9 and leads to endocytosis.
  • Cellular localization
    Membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Antigen identified by monoclonal antibody BB2 antibody
    • BB 2 antibody
    • BB2 antibody
    • CD 54 antibody
    • CD_antigen=CD54 antibody
    • CD54 antibody
    • Cell surface glycoprotein P3.58 antibody
    • Human rhinovirus receptor antibody
    • ICAM 1 antibody
    • ICAM-1 antibody
    • ICAM1 antibody
    • ICAM1_HUMAN antibody
    • intercellular adhesion molecule 1 (CD54), human rhinovirus receptor antibody
    • Intercellular adhesion molecule 1 antibody
    • Major group rhinovirus receptor antibody
    • MALA 2 antibody
    • MALA2 antibody
    • MyD 10 antibody
    • MyD10 antibody
    • P3.58 antibody
    • Surface antigen of activated B cells, BB2 antibody
    see all

Images

  • All lanes : Anti-ICAM1 antibody [MEM-111] (ab2213)

    Lane 1 : TNF-alpha activated HUVEC cells
    Lane 2 : TNF-alpha nonactivated HUVEC cells

    Performed under non-reducing conditions.

    Observed band size: 90 kDa
    why is the actual band size different from the predicted?



    Lower bands represent tubulin as a loading control.

  • Standard Curve for ICAM1 (Analyte: ab82125) dilution range 1pg/ml to 1ug/ml using Capture Antibody Mouse monoclonal [MEM-111] to ICAM1 (ab2213) at 5ug/ml and Detector Antibody Rabbit polyclonal to ICAM1 (Biotin) (ab7815) at 0.5ug/ml
  • ab2213 staining human PBL cells by flow cytometry.  Cells were formaldehyde fixed and gated on B-cells.  The primary antibody was incubated with the sample for 30 minutes at 25°C.  The secondary antibody was ab6810 (a FITC conjugated chicken polyclonal to mouse IgG - H&L).  Both antibodies were used undiluted.

    See Abreview

  • All lanes : Anti-ICAM1 antibody [MEM-111] (ab2213) at 1/3000 dilution

    Lane 1 : Western Markers.
    Lane 2 : Whole cell lysate prepared from human HUVEC cells (untreated) at 100,000 cells.
    Lane 3 : Whole cell lysate prepared from human HUVEC cells (treated with 1 ng/ml TNF-alpha for 24 h) at 100,000 cells.

    Secondary
    All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728)

    Observed band size: 95 kDa why is the actual band size different from the predicted?
    Additional bands at: 40 kDa (possible non-specific binding)

    See Abreview

References

This product has been referenced in:
  • Li Y  et al. Early intervention with mesenchymal stem cells prevents nephropathy in diabetic rats by ameliorating the inflammatory microenvironment. Int J Mol Med 41:2629-2639 (2018). Read more (PubMed: 29484379) »
  • Kotha S  et al. Microvasculature-directed thrombopoiesis in a 3D in vitro marrow microenvironment. PLoS One 13:e0195082 (2018). ICC/IF . Read more (PubMed: 29617409) »
See all 23 Publications for this product

Customer reviews and Q&As

1-10 of 35 Abreviews or Q&A

Abcam has not validated the combination of species/application used in this Abreview.
Application
Blocking
Sample
Streptococcus pneumoniae Cell (Primary Bronchial Epithelial cells)
Specification
Primary Bronchial Epithelial cells

Dr. Selvarani Vimalanathan

Verified customer

Submitted Aug 03 2018

Application
Immunocytochemistry
Sample
Human Cultured Cells (BEAS-2B Epithelial cells)
Permeabilization
Yes - 0.5% saponin
Specification
BEAS-2B Epithelial cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Selvarani Vimalanathan

Verified customer

Submitted Aug 04 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer cell lines total lysate)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
20 µg
Specification
Cancer cell lines total lysate
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 07 2015

Application
Western blot
Loading amount
40 µg
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Sample
Rhesus monkey Cell lysate - whole cell (RF/6A cells from ATCC)
Specification
RF/6A cells from ATCC
Treatment
5.5 mM glucose, 30 mM glucose, 30 mM glucose + 0.1 mM BAPN for 10 days
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 26 2014

Answer

I sorry to hear about the volume of the antibody; we always fill vials 4-5ul extra so quick spin is required to extract the whole volume. I am sure you have given a quick spin to vial before opening it, if it hasn't been done then it will worth to try.

On the other hand I have let our logistic team know about this issue and I can reassure you that they have taken the necessary actions.

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (rat cortex (injured))
Specification
rat cortex (injured)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% triton X-100 in blocking buffer
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Mar 20 2013

Answer

Thank you for your enquiry.

As the Swiss-Prot database highlights, the mature ICAM1 protein is heavily N-Glycosylated. There at least 9 potential site for this type of post-translational modification which can significantly alter the band size. I would advise you to consult these sites for further information:

http://www.uniprot.org/uniprot/P05362

http://www.uniprot.org/uniprot/Q00238

If you need any further assistance in the future, please do not hesitate to contact me.

Read More

Question

Thank you very much,

We used ICAM-1 (Ab, ab2213), but this antibody gives many unspecific bands. I tried it in many different conditions, however, it still gives me the same results.

· Lysis buffer - would be very important to know

We are using special a kit from abcam ab65400 (Plasma membrane protein extraction kit) in which lysis buffer is included.

· Amount of protein loaded - (protein endogenously expressed or transfected cells)

20-30 μg protein per lane.

· Electrophoresis and transfer conditions

The samples were run at 100 and 150 V, one dimensional SDS-PAGE gel was used. For transfering, Protein transfer verifiedusing Ponceau red.

· Blocking conditions

We used 5% and 3% fat free dry milk and also we tried 5% and 3% BSA. Blocking time, once 1 hour and in other run was 30 minutes at 25˚C

· Primary antibody conditions (diluent/dilution, incubation time/temperature)

Concentration or dilution

1/1000 to 1/250 under reducing and non-reducing condition.

· Diluent buffer

1X PBS+0.1% Tween in 5% BSA.

· Incubation time

Once over night at 4C and another 1 hour and 30 minutes at room temperature.

· Secondary antibody conditions (diluent/dilution, incubation time/temperature)

Secondary antibody species gout which reacts against mouse antibodies, the dilution is from 1/10000 to 1/20000 in 1X PBS+0.1% Tween in 5% BSA for one hour at 25˚C.

Wash steps and washing buffer: 1X PBS+0.1% Tween 3 times each one 10 minutes.

· Detection method:

ECL (super signal west pico chemiluminescent)

· Positive control
I used Calnexin, and Na/K ATPase (ab7671) as membrane markerThank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

Read More
Answer

Thank you for your response.

Could you please specify the size of the non-specific bands? Are they lower or higher than the expected size (58 kDa)? Could you attach an image (saved as a jpeg file) to your e-mail so that I can have a look at the bands.

I look forward to hearing from you soon.

Read More

Answer

Thank you for your enquiry.

I can confirm that ab18414 can be stored at 4oC as described on the datasheet. This should not be aliquotted and frozen.

The others I would recommend to aliquot and freeze immediately. Keeping them at 4oC for a few weeks should not significantly affect the staining. However, we would recommend to follow the individual storage instructions provided on the individual datasheets as soon as possible after delivery. This will help to ensure that the antibody remains stable and that we can continue to provide our guarantee.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

Read More

Question
Answer

Thank you for calling us yesterday and for alerting us to the problem you are experiencing with our product. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

As we discussed on the phone, I have attached our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results.

This information will also allow us to investigate this case internally and initiate additional testing where necessary.

I look forward to receiving your reply.

Read More

1-10 of 35 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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