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CD45 Recombinant Rabbit Monoclonal Antibody [PSH13-01] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA751456

CD45 Recombinant Rabbit Monoclonal Antibody [PSH13-01] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IHC-Fr

  • FC

  • IF-Cell

  • IF-Tissue

Reactivity

  • Mouse

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CD45 Recombinant Rabbit Monoclonal Antibody [PSH13-01] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse CD45 aa 1-566.

Species Reactivity

Mouse

Validated Applications

WB, IHC-P, IHC-Fr, FC, IF-Cell, IF-Tissue

Molecular Weight

Predicted band size: 145 kDa

Positive Control

RAW264.7 cell lysate, A20 cell lysate, mouse spleen tissue, mouse stomach tissue, RAW264.7, C57BL/6 mouse spleen cells.

Conjugation

unconjugated

Clone Number

PSH13-01

Product Features

Form

Liquid

Concentration

1mg/ml

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:5,000

  • IHC-P

  • 1:1,000

  • IHC-Fr

  • 1:500

  • FC

  • 1:1,000

  • IF-Cell

  • 1:100

  • IF-Tissue

  • 1:200

Target

Function

CD45, also known as Protein Tyrosine Phosphatase Receptor Type C (PTPRC), is a member of the protein tyrosine phosphatase (PTP) family. It is a type I transmembrane protein expressed on the surface of all differentiated hematopoietic cells, except erythrocytes and plasma cells. CD45 plays a crucial role in regulating T-cell and B-cell antigen receptor signaling, which is essential for immune cell activation and function. CD45 consists of an extracellular domain, a single transmembrane segment, and two tandem intracytoplasmic catalytic domains. While only one of the cytoplasmic domains has intrinsic phosphatase activity, both domains are necessary for substrate recruitment. CD45 activates Src family kinases, such as Lck, which are required for T-cell receptor (TCR) signaling. It also modulates the activity of other kinases, including LYN and FYN, and suppresses JAK kinases, thereby acting as a negative regulator of cytokine receptor signaling. CD45 is involved in various cellular processes, including cell growth, differentiation, and immune response regulation. Dysregulation of CD45 function can lead to severe combined immunodeficiency and is implicated in autoimmune diseases, cancer, and infectious diseases. Additionally, CD45 exists in multiple isoforms, which are used in immunohistochemistry to differentiate between immune cell types and diagnose lymphomas and carcinomas.

Background References

1. Katagiri T, et al. CD45 negatively regulates lyn activity by dephosphorylating both positive and negative regulatory tyrosine residues in immature B cells. J Immunol. 1999, Aug.

2. Holmes N. CD45: all is not yet crystal clear. Immunology. 2006, Feb.

Subcellular Location

Cell membrane, Membrane raft, Synapse.

UNIPROT

SwissProt: P06800 Mouse

Synonyms

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

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of CD45 on different lysates with Rabbit anti-CD45 antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/5,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: C2C12 cell lysate (negative) <br />Lane 3: A20 cell lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 145 kDa<br />Observed band size: 250 kDa<br /><br />Exposure time: 3 minutes; ECL: K1802;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/5,000 dilution was used in primary antibody dilution (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of CD45 on different lysates with Rabbit anti-CD45 antibody (HA751456) at 1/5,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: C2C12 cell lysate (negative)
    Lane 3: A20 cell lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 145 kDa
    Observed band size: 250 kDa

    Exposure time: 3 minutes; ECL: K1802;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA751456) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CD45 antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CD45 antibody (HA751456) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751456) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-CD45 antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/1,000 dilution.<br /><br />The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

    Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-CD45 antibody (HA751456) at 1/1,000 dilution.

    The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA751456) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  • Application: IHC-Fr<br /><br />Species: Mouse<br /><br />Site: spleen<br /><br />Sample: Frozen section<br /><br />Antibody concentration: 1/500<br /><br />Antigen retrieval: Not required

    Application: IHC-Fr

    Species: Mouse

    Site: spleen

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Not required

  • Application: IF-Tissue<br /><br />Species: Mouse<br /><br />Site: spleen<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/200

    Application: IF-Tissue

    Species: Mouse

    Site: spleen

    Sample: Paraffin-embedded section

    Antibody concentration: 1/200

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Immunocytochemistry analysis of RAW264.7 (positive) and C2C12 (negative) labeling CD45 with Rabbit anti-CD45 antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD45 antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Relative expression (RE)

    Immunocytochemistry analysis of RAW264.7 (positive) and C2C12 (negative) labeling CD45 with Rabbit anti-CD45 antibody (HA751456) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD45 antibody (HA751456) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

  • Flow cytometric analysis of C57BL/6 mouse spleen cells labeling CD45.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA751456" style="font-weight: bold;text-decoration: underline;">HA751456</a>, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of C57BL/6 mouse spleen cells labeling CD45.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA751456, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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