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Anti-CD45 antibody [RM1007] (ab281586)

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
Suitable for: IP, Flow Cyt (Intra), WB, IHC-P, ICC
Reacts with: Mouse, Rat, Human

Product name

Anti-CD45 antibody [RM1007]
See all CD45 primary antibodies
Description

Rabbit recombinant multiclonal [RM1007] to CD45
Host species

Rabbit
Tested applications

Suitable for: IP, Flow Cyt (Intra), WB, IHC-P, ICCmore details
Unsuitable for: IHC-Fr
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 corresponding to Mouse CD45 aa 1200-1300.
Database link: P06800

Run BLAST with Run BLAST with
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: Human PBMCs, Jurkat cells, Mouse PBMCs. IP: Jurkat cell.
General notes
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.

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: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS
Concentration information loading...
Purity

Protein A purified
Clonality

Recombinant Multiclonal
Clone number

RM1007
Isotype

IgG

Western blot - Anti-CD45 antibody [RM1007] (ab281586)

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 : 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 (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

Predicted band size: 147 kDa
Observed band size: 180,240 kDa why is the actual band size different from the predicted?

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

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

Negative control: 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] (ab281586)

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] (ab281586)

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] (ab281586)

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] (ab281586)

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] (ab281586)

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] (ab281586)

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 - Anti-CD45 antibody [RM1007] (ab281586)

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 - Anti-CD45 antibody [RM1007] (ab281586)

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 - Anti-CD45 antibody [RM1007] (ab281586)

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] (ab281586)

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] (ab281586)

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