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RecombinantRabMAb

Recombinant Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

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Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Immunocytochemistry/ Immunofluorescence - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Flow Cytometry (Intracellular) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
  • Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23804-95] to CGRP-I+CGRP-II - BSA and Azide free
  • Suitable for: IHC-Fr, WB, Flow Cyt (Intra), IHC-P, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Unconjugated

Overview

  • Product name

    Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free
    See all CGRP-I+CGRP-II primary antibodies

  • Description

    Rabbit monoclonal [EPR23804-95] to CGRP-I+CGRP-II - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-Fr, WB, Flow Cyt (Intra), IHC-P, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: TT whole cell lysate, His-tagged mouse CALC-II recombinant protein and Mouse CALC-I recombinant protein lysates. IHC-P: Mouse spinal cord, Rat spinal cord and Mouse colon tissues. IHC-Fr: Mouse spinal cord and Rat spinal cord. ICC/IF: TT cells. Flow Cyt-intra: TT cells.

  • General notes

    ab283580 is the carrier-free version of ab283568.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab283580 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
IHC-Fr
Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.
Notes
IHC-Fr
Use at an assay dependent concentration.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100.
Application notes
Is unsuitable for IP.

Target

Images

  • Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    All lanes : Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (ab283568) at 1/1000 dilution

    Lane 1 : TT (human thyroid carcinoma epithelial cell), whole cell lysate
    Lane 2 : 293T (human embryonic kidney epithelial cell), whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 4 : Daudi (human Burkitts lymphoma lymphoblast), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 2-4 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Observed band size: 10 kDa why is the actual band size different from the predicted?



    This data was developed using ab283568, the same antibody clone in a different buffer formulation. 

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

    Fresh lysate of TT cell line was used in this blot.

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

    Exposure time: 26 seconds

     

     

  • Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Western blot - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    All lanes : Anti-CGRP-I+CGRP-II antibody [EPR23804-95] (ab283568) at 1/1000 dilution

    Lane 1 : His-tagged mouse CALCB recombinant protein
    Lane 2 : Mouse CALCA recombinant protein at 0.01 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Observed band size: 14 kDa why is the actual band size different from the predicted?



    This data was developed using ab283568, the same antibody clone in a different buffer formulation. 

    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

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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

     

  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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).

     

  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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).

     

  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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).

     

  • Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunohistochemistry (Frozen sections) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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).

     

  • Immunocytochemistry/ Immunofluorescence - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Immunocytochemistry/ Immunofluorescence - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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 Cytometry (Intracellular) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Flow Cytometry (Intracellular) - Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

    This data was developed using ab283568, the same antibody clone in a different buffer formulation.

    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.

  • Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)
    Anti-CGRP-I+CGRP-II antibody [EPR23804-95] - BSA and Azide free (ab283580)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (0)

Publishing research using ab283580? Please let us know so that we can cite the reference in this datasheet.

ab283580 has not yet been referenced specifically in any publications.

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