Product datasheet
Anti-CD18 (phospho S745) antibody [EP1288Y] ab52920
4 Images
Overview
Product name
Anti-CD18 (phospho S745) antibody [EP1288Y]
Description
Rabbit monoclonal [EP1288Y] to CD18 (phospho S745)
Specificity
This antibody only detects CD18 phosphorylated on Serine 745.
Tested applications
WB, IP, IHC-P, ICC/IF, Flow Cyt
Species reactivity
Reacts with: Mouse, Rat, Human
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser745 of human CD18.
Positive control
Jurkat cell lysate or human tonsil tissue. This antibody gave a positive result when used in the
following formaldehyde fixed cell lines: Raw 246.7
General notes
This product is a recombinant rabbit monoclonal antibody.
Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the
following U.S. Patents, No. 5,675,063 and/or 7,429,487.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer
PBS 49%,Sodium azide 0.01%,Glycerol 50%,BSA 0.05%
Purity
Tissue culture supernatant
Clonality
Monoclonal
Clone number
EP1288Y
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab52920 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
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Application
Abreviews
WB
Notes
1/500. Detects a band of approximately 85 kDa (predicted molecular weight: 85
kDa).
IP
1/100.
IHC-P
Use at an assay dependent concentration.
ICC/IF
1/250 - 1/500.
Flow Cyt
1/1000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype
control with this antibody.
Target
Function
Integrin alpha-L/beta-2 is a receptor for ICAM1, ICAM2, ICAM3 and ICAM4. Integrins alphaM/beta-2 and alpha-X/beta-2 are receptors for the iC3b fragment of the third complement
component and for fibrinogen. Integrin alpha-X/beta-2 recognizes the sequence G-P-R in
fibrinogen alpha-chain. Integrin alpha-M/beta-2 recognizes P1 and P2 peptides of fibrinogen
gamma chain. Integrin alpha-M/beta-2 is also a receptor for factor X. Integrin alpha-D/beta-2 is a
receptor for ICAM3 and VCAM1.
Involvement in disease
Defects in ITGB2 are the cause of leukocyte adhesion deficiency type 1 (LAD1) [MIM:116920].
LAD1 patients have recurrent bacterial infections and their leukocytes are deficient in a wide
range of adhesion-dependent functions.
Sequence similarities
Belongs to the integrin beta chain family.
Contains 1 VWFA domain.
Post-translational
modifications
Both Ser-745 and Ser-756 become phosphorylated when T-cells are exposed to phorbol esters.
Phosphorylation on Thr-758 (but not on Ser-756) allows interaction with 14-3-3 proteins.
Cellular localization
Membrane.
Anti-CD18 (phospho S745) antibody [EP1288Y] images
2
ICC/IF image of ab52920 stained Raw 246.7
cells. The cells were 4% formaldehyde fixed
(10 min) and then incubated in 1%BSA / 10%
normal goat serum / 0.3M glycine in 0.1%
PBS-Tween for 1h to permeabilise the cells
and block non-specific protein-protein
interactions. The cells were then incubated
with the antibody ab52920 at 1/1000 dilution
Immunocytochemistry/ Immunofluorescence Anti-CD18 (phospho S745) antibody [EP1288Y]
(ab52920)
overnight at +4°C. The secondary antibody
(green) was DyLight® 488 goat anti- rabbit
(ab96899) IgG (H+L) used at a 1/250 dilution
for 1h. Alexa Fluor® 594 WGA was used to
label plasma membranes (red) at a 1/200
dilution for 1h. DAPI was used to stain the cell
nuclei (blue) at a concentration of 1.43µM.
All lanes : Anti-CD18 (phospho S745)
antibody [EP1288Y] (ab52920) at 1/500
dilution
Lane 1 : Jurkat cell lysate, untreated
Lane 2 : Jurkat cell lysate, treated with AP.
Lysates/proteins at 10 µg per lane.
Secondary
Goat anti-rabbit HRP labeled at 1/2000
Western blot - CD18 (phospho S745) antibody
dilution
[EP1288Y] (ab52920)
Predicted band size : 85 kDa
Observed band size : 85 kDa
Ab52920 (1:250) staining human CD18 in
human tonsil tissue by immunohistochemistry
using paraffin embedded tissue.Ab52920
(1:250) staining human CD18 in human tonsil
tissue by immunohistochemistry using paraffin
embedded tissue.
Immunohistochemistry (Paraffin-embedded
sections) - CD18 (phospho S745) antibody
[EP1288Y] (ab52920)
3
Human peripheral blood lymphocytes stained
with ab52920 (red line). Human whole blood
was processed using a modified protocol
based on Chow et al, 2005 (PMID:
16080188). In brief, human whole blood was
fixed in 4% formaldehyde (methanol-free) for
10 min at 22°C. Red blood cells were then
lyzed by the addition of Triton X-100 (final
concentration - 0.1%) for 15 min at 37°C. For
Flow Cytometry - Anti-CD18 (phospho S745)
antibody [EP1288Y] (ab52920)
experimentation, cells were treated with 50%
methanol (-20°C) for 15 min at 4°C. Cells
were then incubated with the antibody
(ab52920, 1/1000 dilution) for 30 min at 4°C.
The secondary antibody used was Alexa
Fluor® 488 goat anti-rabbit IgG (H&L)
(ab150077) at 1/2000 dilution for 30 min at
4°C. Isotype control antibody (black line) was
rabbit IgG (monoclonal) (0.1μg/1x106 cells)
used under the same conditions. Unlabelled
sample (blue line) was also used as a control.
Acquisition of >30,000 total events were
collected using a 20mW Argon ion laser
(488nm) and 525/30 bandpass filter. Gating
strategy - peripheral blood lymphocytes.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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