重组Anti-CGRP-I+CGRP-II抗体[EPR23804-95]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized TT cells labelling CGRP-I+CGRP-II with ab283568 at 1/100 (6.39 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Confocal image showing cytoplasmic staining in TT cell line, and no staining in 293T cell line is observed.

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell) (Left) / TT (Human thyroid carcinoma epithelial cell) (Right) cells labelling CGRP-I+CGRP-II with ab283568 at 1/5000 dilution (0.01ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat F(ab')2 Anti-Rabbit IgG(DyLight® 488, ab98507) at 1/500 dilution was used as the secondary antibody.

Negative control : 293T.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of paraffin-embedded Mouse spinal cord tissue labelling CGRP-I+CGRP-II with ab283568 at 1/1000 (0.639 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse central termination sensory neurons (PMID : 29634489). The section was incubated with ab283568 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat spinal cord tissue labeling CGRP-I+CGRP-II with ab283568 at 1/100 (6.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on the dorsal horn of rat spinal cord is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of paraffin-embedded Rat skin tissue labelling CGRP-I+CGRP-II with ab283568 at 1/1000 (0.639 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The section was incubated with ab283568 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Negative control : no staining on rat skin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse skin tissue labeling CGRP-I+CGRP-II with ab283568 at 1/100 (6.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution (Green). The nuclear counterstain was DAPI (Blue).

Negative control : No staining on mouse skin is observed.

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of paraffin-embedded Rat spinal cord tissue labelling CGRP-I+CGRP-II with ab283568 at 1/1000 (0.639 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat central termination sensory neurons. The section was incubated with ab283568 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat skin tissue labeling CGRP-I+CGRP-II with ab283568 at 1/100 (6.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). The nuclear counterstain was DAPI (Blue).

Negative control : No staining on rat skin is observed.

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labelling CGRP-I+CGRP-II with ab283568 at 1/1000 (0.639 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the muscularis nerve fibres of mouse colon. The section was incubated with ab283568 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labelling CGRP-I+CGRP-II with ab283568 at 1/1000 (0.639 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The section was incubated with ab283568 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

Negative control : no staining on mouse skin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunofluorescent analysis of 4% Paraformaldehyde fixed rat DRG primary neurons permeabilized using 0.1% Triton X-100 labelling CGRP-I+CGRP-II with ab283568 (green) followed by ab150081 AlexaFluor^®488 preadsorbed Goat anti-Rabbit secondary. ab78078 anti-beta III Tubulin (mouse mAb) was used as counterstain (magenta) with ab150120 AlexaFluor^®594 Goat anti-Mouse as secondary. Nuclear DNA was labelled with DAPI (shown in blue). Image showing cytoplasmic staining in rat DRG primary neurons was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunofluorescent analysis of 4% Paraformaldehyde fixed mouse DRG primary neurons permeabilized using 0.1% Triton X-100 labelling CGRP-I+CGRP-II with ab283568 (green) followed by ab150081 AlexaFluor^®488 preadsorbed Goat anti-Rabbit secondary. ab78078 anti-beta III Tubulin (mouse mAb) was used as counterstain (magenta) with ab150120 AlexaFluor^®594 Goat anti-Mouse as secondary. Nuclear DNA was labelled with DAPI (shown in blue). Image showing cytoplasmic staining in mouse DRG primary neurons was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spinal cord tissue labeling CGRP-I+CGRP-II with ab283568 at 1/100 (6.39 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution (Green). Positive staining on the dorsal horn of mouse spinal cord is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• WB

Lab

Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Blocking and diluting buffer and concentration : 5% NFDM/TBSTFresh lysate of TT cell line was used in this blot.

Negative control : 293T, Hela, Daudi (HPA database)

Exposure time : 26 seconds

All lanes:

Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (ab283568) at 1/1000 dilution

Lane 1:

TT (human thyroid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

Daudi (human Burkitt's lymphoma lymphoblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 10 kDa

false

• WB

Lab

Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (AB283568)

Blocking and diluting buffer and concentration : 5% NFDM/TBST

This antibody has cross-reaction with mouse CALCB.

The rec proteins CALCB and CLACA were made in-house.

CALCB recombinant protein was made in-house and expressed from the E.coli expression system.

CLACA recombinant protein was expressed from a mammalian - HEK-293 expression system.

Exposure time : 1 second

All lanes:

Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (ab283568) at 1/1000 dilution

Lane 1:

His-tagged mouse CALCB recombinant protein at 0.01 µg

Lane 2:

Mouse CALCA recombinant protein at 0.01 µg

Secondary

Lanes 1 - 2:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 1 - 2:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 14 kDa

false