Anti-CD75 antibody [LN1] ab77676 Product datasheet 3 Images Overview

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Product datasheet
Anti-CD75 antibody [LN1] ab77676
3 Images
Overview
Product name
Anti-CD75 antibody [LN1]
Description
Mouse monoclonal [LN1] to CD75
Specificity
This antibody is shown to be a reliable antibody for ascribing a B-cell phenotype in known
lymphoid tissues. We have data to indicate that this antibody may not cross react with Rat.
However, this has not been conclusively tested and expression levels may vary in certain cell
lines/tissues.
Tested applications
IHC-P, Flow Cyt, ICC/IF
Species reactivity
Reacts with: Human
Immunogen
Tissue/ cell preparation - Nuclei from pokeweed mitogen-stimulated peripheral blood
lymphocytes.
Positive control
Lymph node or human tonsil.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw
cycles.
Storage buffer
Preservative: None
Constituents: 10mM PBS, pH 7.4
Purity
Ammonium Sulphate Precipitation
Clonality
Monoclonal
Clone number
LN1
Isotype
IgM
Light chain type
kappa
Applications
Our Abpromise guarantee covers the use of ab77676 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
IHC-P
Notes
1/25 - 1/50. Staining of formalin-fixed tissues requires boiling tissue sections in
10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 min.
Use 1µg for 106 cells. ab18400-Mouse monoclonal IgM, is suitable for use as an
Flow Cyt
isotype control with this antibody.
ICC/IF
Use at an assay dependent concentration.
Target
Function
Transfers sialic acid from the donor of substrate CMP-sialic acid to galactose containing
acceptor substrates.
Pathway
Protein modification; protein glycosylation.
Sequence similarities
Belongs to the glycosyltransferase 29 family.
Post-translational
modifications
The soluble form derives from the membrane form by proteolytic processing.
The HB-6, CDW75, and CD76 differentiation antigens are cell-surface carbohydrate
determinants generated by this enzyme.
Cellular localization
Golgi apparatus > Golgi stack membrane. Secreted. Membrane-bound form in trans cisternae of
Golgi. Secreted into the body fluid.
Anti-CD75 antibody [LN1] images
Formalin-fixed, paraffin-embedded human
tonsil stained with ab77676 at 4-8ug/ml, using
a peroxidase-conjugate secondary and AEC
chromogen. Note cell membrane and
cytoplasmic staining of B cells in germinal
center.
Immunohistochemistry (Formalin/PFA-fixed
paraffin-embedded sections) - CD75 antibody
[LN1] - BSA and Azide free (ab77676)
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ICC/IF image of ab77676 stained HeLa cells.
The cells were 100% methanol fixed (5 min)
and then incubated in 1%BSA / 10% normal
goat serum / 0.3M glycine in 0.1% PBSTween for 1h to permeabilise the cells and
block non-specific protein-protein
interactions. The cells were then incubated
with the antibody (ab77676, 10µg/ml)
overnight at +4°C. The secondary antibody
Immunocytochemistry/ Immunofluorescence-
(green) was ab96879, DyLight® 488 goat
CD75 antibody [LN1] - BSA and Azide
anti-mouse IgG (H+L) used at a 1/250 dilution
free(ab77676)
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.
Overlay histogram showing Ramos cells
stained with ab77676 (red line). The cells
were fixed with 4% paraformaldehyde (10
min) and then permeabilized with 0.1% PBSTween 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
Flow Cytometry-Anti-CD75 antibody [LN1] - BSA
antibody (ab77676, 1µg/1x106 cells) for 30
and Azide free(ab77676)
min at 22ºC. The secondary antibody used
was DyLight® 488 goat anti-mouse IgG (H+L)
(ab96879) at 1/500 dilution for 30 min at
22ºC. Isotype control antibody (black line) was
mouse IgM [ICIGM] (ab91545, 2µg/1x106
cells) used under the same conditions.
Acquisition of >5,000 events was performed.
This antibody gave a positive signal in Ramos
cells fixed with 80% methanol (5
min)/permeabilized with 0.1% PBS-Tween for
20 min used under the same conditions.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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Replacement or refund for products not performing as stated on the datasheet
Valid for 12 months from date of delivery
Response to your inquiry within 24 hours
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