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RecombinantRabMAb

Recombinant Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

Reviews (1) Submit a question
Western blot - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunocytochemistry - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Immunoprecipitation - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
  • Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit recombinant multiclonal [RM1007] to CD45 - BSA and Azide free
  • Suitable for: IHC-P, ICC, Flow Cyt (Intra), IP, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

Overview

  • Product name

    Anti-CD45 antibody [RM1007] - BSA and Azide free
    See all CD45 primary antibodies

  • Description

    Rabbit recombinant multiclonal [RM1007] to CD45 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC, Flow Cyt (Intra), IP, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    This product was produced with the following immunogens:
    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Jurkat, Mouse thymus, Rat thymus lysates. IHC-P: Human tonsil, spleen, colon cancer, Mouse spleen and Rat spleen tissues. ICC: Jurkat cells. Flow Cyt (intra): Human PBMCs, Jurkat cells, Mouse PBMCs. IP: Jurkat cell.

  • General notes

    ab282747 is the carrier free version of ab281586.

    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.

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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab282747 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-P (1)
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 147 kDa.
Notes
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 147 kDa.

Target

  • Function

    Protein tyrosine-protein phosphatase required for T-cell activation through the antigen receptor. Acts as a positive regulator of T-cell coactivation upon binding to DPP4. The first PTPase domain has enzymatic activity, while the second one seems to affect the substrate specificity of the first one. Upon T-cell activation, recruits and dephosphorylates SKAP1 and FYN.

  • Involvement in disease

    Defects in PTPRC are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)B(+)NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development.
    Genetic variations in PTPRC are involved in multiple sclerosis susceptibility (MS) [MIM:126200]. MS is a neurodegenerative disorder characterized by the gradual accumulation of focal plaques of demyelination particularly in the periventricular areas of the brain. Peripheral nerves are not affected. Onset usually in third or fourth decade with intermittent progression over an extended period. The cause is still uncertain.

  • Sequence similarities

    Belongs to the protein-tyrosine phosphatase family. Receptor class 1/6 subfamily.
    Contains 2 fibronectin type-III domains.
    Contains 2 tyrosine-protein phosphatase domains.

  • Domain

    The first PTPase domain interacts with SKAP1.

  • Post-translational
    modifications

    Heavily N- and O-glycosylated.

  • Cellular localization

    Membrane. Membrane raft. Colocalized with DPP4 in membrane rafts.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • B220 antibody
    • CD 45 antibody
    • CD45 antibody
    • CD45 antigen antibody
    • CD45R antibody
    • GP180 antibody
    • L-CA antibody
    • LCA antibody
    • Leukocyte common antigen antibody
    • loc antibody
    • Ly-5 antibody
    • LY5 antibody
    • Ly5, homolog of antibody
    • Lyt-4 antibody
    • OTTHUMP00000033813 antibody
    • OTTHUMP00000033816 antibody
    • OTTHUMP00000033817 antibody
    • OTTHUMP00000038574 antibody
    • Protein tyrosine phosphatase receptor type c polypeptide antibody
    • Protein tyrosine phosphatase, receptor type C antibody
    • protein tyrosine phosphatase, receptor type, C antibody
    • Protein tyrosine phosphatase, receptor type, c polypeptide antibody
    • Ptprc antibody
    • PTPRC_HUMAN antibody
    • Receptor-type tyrosine-protein phosphatase C antibody
    • T200 antibody
    • T200 glycoprotein antibody
    • T200 leukocyte common antigen antibody
    see all

Images

  • Western blot - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Western blot - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    All lanes : Anti-CD45 antibody [RM1007] (ab281586) at 1/1000 dilution

    Lane 1 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
    Lane 2 : HEK-293T ( human embryonic kidney epithelial cell) whole cell lysate
    Lane 3 : Mouse thymus tissue lysate
    Lane 4 : Rat thymus tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 147 kDa



    This data was produced using ab281586, the same clone in a different formulation.

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

    The MW observed is consistent with the literature (PMID: 14715639).

    Negative control: HEK-293T (PMID: 22978632).

    This blot was developed using a higher sensitivity ECL substrate.

    Exposure time: 3 minutes.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labelling CD45 with ab281586 at 1/4000 (0.138 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human tonsil. The section was incubated with ab281586 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded Human spleen tissue labelling CD45 with ab281586 at 1/4000 (0.138 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on human spleen. The section was incubated with ab281586 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labelling CD45 with ab281586 at 1/4000 (0.138 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stromal cells of human colon cancer (PMID: 30713795). The section was incubated with ab281586 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labelling CD45 with ab281586 at 1/2000 (0.275 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse spleen. The section was incubated with ab281586 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labelling CD45 with ab281586 at 1/2000 (0.275 ug/ml) dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat spleen. The section was incubated with ab281586 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunocytochemistry - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunocytochemistry - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labelling CD45 with ab281586 at 1/50 (11.0 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green) Confocal image showing membranouse staining in Jurkat cells and no staining in 293T cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human peripheral blood mononuclear cell (PBMC) cells labelling CD45 with ab281586 at 1/500 dilution (0.1ug). Right compared with a Rabbit monoclonal IgG (ab172730)/ Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (Human embryonic kidney epithelial cell, Left) / Jurkat (Human T cell leukemia T lymphocyte, Right) cells labelling CD45 with ab281586 at 1/500 dilution (0.1ug) (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 anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Negative control: 293T. (PMID: 16005866)

  • Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Flow Cytometry (Intracellular) - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse peripheral blood mononuclear cell (PBMC) cells labelling CD45 with ab281586 at 1/50 dilution (1ug). Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Immunoprecipitation - Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

    This data was produced using ab281586, the same clone in a different formulation.

    CD45 was immunoprecipitated from 0.35 mg Jurkat (human T cell leukemia T lymphocyte), whole cell lysate 10 ug with ab281586 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using 281586 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: Jurkat (human T cell leukemia T lymphocyte), whole cell lysate 10 ug

    Lane 2: ab281586 IP in Jurkat whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of abx in Jurkat whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

  • Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)
    Anti-CD45 antibody [RM1007] - BSA and Azide free (ab282747)

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 ab282747? Please let us know so that we can cite the reference in this datasheet.

ab282747 has not yet been referenced specifically in any publications.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD45 antibody [RM1007] - BSA and Azide free

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH 6 & TRIS pH 9
Permeabilization
Yes - 0.3% Triton X-100
Specification
Tonsil
Blocking step
Donkey Serum 10% + 3% BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Dylan Windell

Verified customer

Submitted May 05 2022

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