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CD19 Recombinant Rabbit Monoclonal Antibody [PSH03-94] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750919

CD19 Recombinant Rabbit Monoclonal Antibody [PSH03-94] - BSA and Azide free

Application

  • WB

  • IHC-P

  • IF-Tissue

  • IP

  • IHC-Fr

  • FC

  • IF-Cell

Reactivity

  • Mouse

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

CD19 Recombinant Rabbit Monoclonal Antibody [PSH03-94] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse CD19 protein.

Species Reactivity

Mouse

Validated Applications

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

Molecular Weight

Predicted band size: 60 kDa

Positive Control

Mouse spleen tissue lysates, mouse lymph node tissue, mouse spleen tissue, mouse spleen cells.

Conjugation

unconjugated

Clone Number

PSH03-94

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

  • 1:1,000

  • IHC-P

  • 1:500

  • IF-Tissue

  • 1:200

  • IP

  • 1-2μg/sample

  • IHC-Fr

  • 1:500

  • FC

  • 1:1,000

  • IF-Cell

  • 1:200

Target

Function

B-lymphocyte antigen CD19, also known as CD19 molecule (Cluster of Differentiation 19), B-Lymphocyte Surface Antigen B4, T-Cell Surface Antigen Leu-12 and CVID3 is a transmembrane protein that in humans is encoded by the gene CD19. In humans, CD19 is expressed in all B lineage cells. Contrary to some early doubts, human plasma cells do express CD19, as confirmed by others. CD19 plays two major roles in human B cells: on the one hand, it acts as an adaptor protein to recruit cytoplasmic signaling proteins to the membrane; on the other, it works within the CD19/CD21 complex to decrease the threshold for B cell receptor signaling pathways. Due to its presence on all B cells, it is a biomarker for B lymphocyte development, lymphoma diagnosis and can be utilized as a target for leukemia immunotherapies.

Background References

1. Mougiakakos D et al. CD19-Targeted CAR T Cells in Refractory Systemic Lupus Erythematosus. N Engl J Med. 2021 Aug

2. Jin X et al. Therapeutic efficacy of anti-CD19 CAR-T cells in a mouse model of systemic lupus erythematosus. Cell Mol Immunol. 2021 Aug

Subcellular Location

Cell membrane, Membrane raft.

UNIPROT

SwissProt: P25918 Mouse

Synonyms

Antibody deficiency due to defect in CD19 antibody

Antibody deficiency due to defect in CD19, included antibody

AW495831 antibody

B lymphocyte antigen CD19 antibody

B lymphocyte surface antigen B4 antibody

B-lymphocyte antigen CD19 antibody

B-lymphocyte surface antigen B4 antibody

B4 antibody

CD19 antibody

CD19 antigen antibody

Expand

Images

  • Western blot analysis of CD19 on mouse spleen tissue lysates with Rabbit anti-CD19 antibody (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/1,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 60 kDa<br />Observed band size: 60-120 kDa<br /><br />Exposure time: 30 seconds;<br /><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/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/1,000 dilution was used in 5% NFDM/TBST 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.

    Western blot analysis of CD19 on mouse spleen tissue lysates with Rabbit anti-CD19 antibody (HA750919) at 1/1,000 dilution.

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 60 kDa
    Observed band size: 60-120 kDa

    Exposure time: 30 seconds;

    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 (HA750919) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • 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: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

    Application: IHC-Fr

    Species: Mouse

    Site: spleen

    Sample: Frozen section

    Antibody concentration: 1/500

    Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

  • Immunohistochemical analysis of paraffin-embedded mouse lymph node tissue with Rabbit anti-CD19 antibody (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/500 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/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/500 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 lymph node tissue with Rabbit anti-CD19 antibody (HA750919) at 1/500 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 (HA750919) at 1/500 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-CD19 antibody (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/500 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/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/500 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-CD19 antibody (HA750919) at 1/500 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 (HA750919) at 1/500 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: 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 />Application: IF-Tissue<br /><br />Species: Mouse<br /><br />Site: liver (negative)<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/200

    ☑ Relative expression (RE)

    Application: IF-Tissue

    Species: Mouse

    Site: liver (negative)

    Sample: Paraffin-embedded section

    Antibody concentration: 1/200

  • CD19 was immunoprecipitated in 0.2mg mouse spleen tissue lysate with <a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a> at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.<br /><br />Lane 1: Mouse spleen tissue lysate (input)<br />Lane 2: <a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a> IP in mouse spleen tissue lysate<br />Lane 3: Rabbit IgG instead of <a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a> in mouse spleen tissue lysate<br /><br />Blocking/Dilution buffer: 5% NFDM/TBST<br />Exposure time: 17 seconds; ECL: <a href="/products/K1802" style="font-weight: bold;text-decoration: underline;">K1802</a>

    CD19 was immunoprecipitated in 0.2mg mouse spleen tissue lysate with HA750919 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750919 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: Mouse spleen tissue lysate (input)
    Lane 2: HA750919 IP in mouse spleen tissue lysate
    Lane 3: Rabbit IgG instead of HA750919 in mouse spleen tissue lysate

    Blocking/Dilution buffer: 5% NFDM/TBST
    Exposure time: 17 seconds; ECL: K1802

  • Flow cytometric analysis of mouse spleen cells labeling CD19 (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) and CD3-PE.<br /><br />Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>, 1/1,000). 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℃.

    Flow cytometric analysis of mouse spleen cells labeling CD19 (HA750919) and CD3-PE.

    Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA750919, 1/1,000). 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℃.

  • Immunocytochemistry analysis of mouse spleen cells labeling CD19 with Rabbit anti-CD19 antibody (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/200 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde 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-CD19 antibody (<a href="/products/HA750919" style="font-weight: bold;text-decoration: underline;">HA750919</a>) at 1/200 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.

    Immunocytochemistry analysis of mouse spleen cells labeling CD19 with Rabbit anti-CD19 antibody (HA750919) at 1/200 dilution.

    Cells were fixed in 4% paraformaldehyde 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-CD19 antibody (HA750919) at 1/200 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.

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