Product datasheet
Anti-CRMP1 antibody [EP14521] ab199722
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Overview
Product name
Anti-CRMP1 antibody [EP14521]
Description
Rabbit monoclonal [EP14521] to CRMP1
Tested applications
Flow Cyt, IHC-P, WB, ICC/IF, IP
Species reactivity
Reacts with: Mouse, Rat, Human
Immunogen
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within
Human CRMP1 aa 500 to the C-terminus. The exact sequence is proprietary.
Database link: Q14194
Positive control
WB: Human fetal brain lysate; Mouse brain lysate; SH-SY5Y and C6 cell lysates. IHC-P: Human,
mouse and rat cerebral cortex tissues. ICC/IF: U-87 MG cells. Flow Cyt: SH-SY5Y cells. IP:
Human fetal brain whole cell lysate.
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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long
term. Avoid freeze / thaw cycle.
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Purity
Protein A purified
Clonality
Monoclonal
Clone number
EP14521
Isotype
IgG
Applications
Our Abpromise guarantee covers the use of ab199722 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
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Application
Abreviews
Notes
Flow Cyt
1/20.
IHC-P
1/8000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0
before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 64, 80 kDa (predicted molecular
weight: 62 kDa).
ICC/IF
1/500.
IP
1/30.
Target
Function
Necessary for signaling by class 3 semaphorins and subsequent remodeling of the cytoskeleton.
Plays a role in axon guidance, invasive growth and cell migration.
Tissue specificity
Brain.
Sequence similarities
Belongs to the DHOase family. Hydantoinase/dihydropyrimidinase subfamily.
Cellular localization
Cytoplasm. Cytoplasm > cytoskeleton > centrosome. Cytoplasm > cytoskeleton > spindle.
Associated with centrosomes and the mitotic spindle during metaphase.
Anti-CRMP1 antibody [EP14521] images
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Immunofluorescent analysis of 4%
paraformaldehyde-fixed, 0.1% Triton X-100
permeabilized U-87 MG (Human
glioblastoma-astrocytoma epithelial cell line)
and Jurkat (Human T cell leukemia cells from
peripheral blood) cells labeling CRMP1 with
ab199722 at 1/500 dilution, followed by Goat
anti-rabbit IgG (Alexa Fluor® 488)
(ab150077) secondary antibody at 1/500
dilution (green). Cytoplasm and nuclear
staining on U-87 MG cell line is observed.
Immunocytochemistry/ Immunofluorescence -
Negative expression in Jurkat cells. The
Anti-CRMP1 antibody [EP14521] (ab199722)
nuclear counterstain is DAPI (blue). Tubulin is
detected with ab7291 (anti-Tubulin mouse
mAb) at 1/1000 dilution and ab150120
(AlexaFluor®594 Goat anti-Mouse
secondary) at 1/500 dilution (red).
The negative controls are as follows:-ve control 1: ab199722 at 1/500 dilution
followed by ab150120 (AlexaFluor®594 Goat
anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse
mAb) at 1/1000 dilution followed by
ab150077 (Alexa Fluor®488 Goat AntiRabbit IgG H&L) at 1/500 dilution.
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All lanes : Anti-CRMP1 antibody [EP14521]
(ab199722) at 1/5000 dilution
Lane 1 : Human fetal brain lysate
Lane 2 : SH-SY5Y (Human neuroblastoma
from bone marrow cells) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase
Western blot - Anti-CRMP1 antibody [EP14521]
conjugated at 1/1000 dilution
(ab199722)
Predicted band size : 62 kDa
Observed band size : 64,80 kDa
Exposure time : 15 seconds
Blocking/Dilution buffer: 5% NFDM/TBST.
Based on the sequence analysis, ab199722
recognizes two isoforms with the predicted
MWs of 62 kDa and 74 kDa, respectively.
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All lanes : Anti-CRMP1 antibody [EP14521]
(ab199722) at 1/1000 dilution
Lane 1 : Mouse brain lysate
Lane 2 : C6 (Rat glial tumor cells) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase
conjugated at 1/1000 dilution
Western blot - Anti-CRMP1 antibody [EP14521]
(ab199722)
Predicted band size : 62 kDa
Observed band size : 64 kDa
Exposure time : 30 seconds
Blocking buffer: 5% NFDM/TBST.
Dilution buffer: 5% NFDM /TBST or 1%BSA
/TBST.
Based on the sequence analysis, CRMP1
does not have similar isoforms within Mouse
& Rat, therefore only a single band at
approximately 62kDa is seen in Mouse and
Rat lysates within western blot.
Immunohistochemical analysis of paraffinembedded Human cerebral cortex tissue
labeling CRMP1 with ab199722 at 1/8000
dilution, followed by Goat Anti-Rabbit IgG
H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm and nucleus staining on Human
cerebral cortex is observed. Counter stained
with Hematoxylin.
Secondary antibody only control: Used PBS
instead of primary antibody, secondary
Immunohistochemistry (Formalin/PFA-fixed
antibody is Goat Anti-Rabbit IgG H&L (HRP)
paraffin-embedded sections) - Anti-CRMP1
(ab97051) at 1/500 dilution.
antibody [EP14521] (ab199722)
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Immunohistochemical analysis of paraffinembedded Mouse cerebral cortex tissue
labeling CRMP1 with ab199722 at 1/8000
dilution, followed by Goat Anti-Rabbit IgG
H&L (HRP) (ab97051) at 1/500 dilution.
Cytoplasm staining on mouse cerebral cortex
is observed. Counter stained with
Hematoxylin.
Secondary antibody only control: Used PBS
instead of primary antibody, secondary
Immunohistochemistry (Formalin/PFA-fixed
antibody is Goat Anti-Rabbit IgG H&L (HRP)
paraffin-embedded sections) - Anti-CRMP1
(ab97051) at 1/500 dilution.
antibody [EP14521] (ab199722)
Immunohistochemical analysis of paraffinembedded Rat cerebral cortex tissue labeling
CRMP1 with ab199722 at 1/8000 dilution,
followed by Goat Anti-Rabbit IgG H&L (HRP)
(ab97051) at 1/500 dilution. Cytoplasm
staining on rat cerebral cortex is observed.
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS
instead of primary antibody, secondary
antibody is Goat Anti-Rabbit IgG H&L (HRP)
Immunohistochemistry (Formalin/PFA-fixed
(ab97051) at 1/500 dilution.
paraffin-embedded sections) - Anti-CRMP1
antibody [EP14521] (ab199722)
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Immunohistochemical analysis of paraffinembedded Human liver tissue labeling
CRMP1 with ab199722 at 1/8000 dilution,
followed by Goat Anti-Rabbit IgG H&L (HRP)
(ab97051) at 1/500 dilution. No staining on
Human liver tissue is observed. Counter
stained with Hematoxylin.
Secondary antibody only control: Used PBS
instead of primary antibody, secondary
antibody is Goat Anti-Rabbit IgG H&L (HRP)
Immunohistochemistry (Formalin/PFA-fixed
(ab97051) at 1/500 dilution.
paraffin-embedded sections) - Anti-CRMP1
antibody [EP14521] (ab199722)
Flow cytometric analysis of 2%
paraformaldehyde-fixed SH-SY5Y (Human
neuroblastoma from bone marrow cells) cells
labeling CRMP1 with ab199722 at 1/20
dilution (red) compared with a rabbit
monoclonal IgG isotype control
(ab172730; black) and an unlabelled control
(cells without incubation with primary antibody
and secondary antibody; blue). Goat anti
Flow Cytometry - Anti-CRMP1 antibody
[EP14521] (ab199722)
rabbit IgG (FITC) at 1/150 dilution was used
as the secondary antibody.
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CRMP1 was immunoprecipitated from 1mg
of Human fetal brain whole cell lysate with
ab199722 at 1/30 dilution. Western blot was
performed from the immunoprecipitate using
ab199722 at 1/1000 dilution. Anti-Rabbit IgG
(HRP), specific to the non-reduced form of
IgG, was used as secondary antibody at
1/1500.
Lane 1: Human fetal brain whole cell lysate
10ug (Input). Lane 2: ab199722 IP in Human
Immunoprecipitation - Anti-CRMP1 antibody
fetal brain whole cell lysate. Lane 3: Rabbit
[EP14521] (ab199722)
monoclonal IgG (ab172730) instead of
ab199722 in Human fetal brain whole cell
lysate.
Blocking and dilution buffer and
concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
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