CD19 Recombinant Rabbit Monoclonal Antibody [PSH03-94]

Western blot analysis of CD19 on mouse spleen tissue lysates with Rabbit anti-CD19 antibody (HA722073) 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 (HA722073) 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

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 (HA722073) 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 (HA722073) 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 (HA722073) 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 (HA722073) 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

Species: Mouse

Site: spleen

Sample: Paraffin-embedded section

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 HA722073 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722073 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: HA722073 IP in mouse spleen tissue lysate
Lane 3: Rabbit IgG instead of HA722073 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 (HA722073) and CD3-PE.

Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA722073, 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 (HA722073) 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 (HA722073) 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.

Fluorescence multiplex immunohistochemical analysis of mouse lymph nodes (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-TCF7 (HA723582, white), anti-CD4 (HA722966, red) and anti-CD19 (HA722073, Yellow) on lymph nodes. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in three rounds of staining: in the order of HA723582 (1/200 dilution), HA722966 (1/500 dilution) and HA722073 (1/500 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.