Anti-CD19 antibody [RM1166]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling CD19 with ab317335 at 1/200 (2.585 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on human cardiac muscle. The section was incubated with ab317335 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD19 with ab317335 at 1/200 (2.585 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil. The section was incubated with ab317335 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling CD19 with ab317335 at 1/200 (2.585 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human spleen. The section was incubated with ab317335 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [RM1166] (AB317335)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling CD19 with ab317335 at 1/100 (5.17 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in Raji cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : Jurkat (PMID : 29928472).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IP

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Immunoprecipitation - Anti-CD19 antibody [RM1166] (AB317335)

CD19 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab317335 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317335 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2 : ab317335 IP in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317335 in Raji whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CD19 antibody [RM1166] (ab317335) at 1/30 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate

Lane 2:

ab317335 IP in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 41s

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen (fresh frozen) tissue labeling CD19 with ab317335 at 1/50 (10.34 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Confocal image showing positive staining on mouse spleen. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317335 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse heart (fresh frozen) tissue labeling CD19 with ab317335 at 1/50 (10.34 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green).

Negative control : confocal image showing no staining on mouse heart. The nuclear counterstain was DAPI (Blue). The section was incubated with ab317335 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbedat 1/1000 2 ug/mL dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD19 with ab317335 at 1/500 (1.034 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab317335 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD19 antibody [RM1166] (AB317335)

Immunohistochemical analysis of paraffin-embedded Mouse cardiac tissue labeling CD19 with ab317335 at 1/500 (1.034 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : no staining on mouse cardiac muscle. The section was incubated with ab317335 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD19 antibody [RM1166] (AB317335)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Tween-20 permeabilized WEHI-231(mouse B cell lymphoma B lymphocyte) cells labelling CD19 with ab317335 at 1/100 (5.17 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in WEHI-231cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : CTLL-2.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-CD19 antibody [RM1166] (AB317335)

CD19 was immunoprecipitated from 0.35 mg WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate with ab317335 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317335 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate
Lane 2 : ab317335 IP in WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317335 in WEHI-231 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CD19 antibody [RM1166] (ab317335) at 1/30 dilution

Lane 1:

WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate

Lane 2:

ab317335 IP in WEHI-231 (mouse B cell lymphoma B lymphocyte) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 84s

• WB

Supplier Data

Western blot - Anti-CD19 antibody [RM1166] (AB317335)

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

All lanes:

Western blot - Anti-CD19 antibody [RM1166] (ab317335) at 1/1000 dilution

Lane 1:

Untreated Raji whole cell lysate at 15 µg

Lane 2:

Raji whole cell lysate treated with Peptide:N-glycosidase F (PNGase F) at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 100 kDa,60-75 kDa

false

Exposure time: 103s

• WB

Supplier Data

Western blot - Anti-CD19 antibody [RM1166] (AB317335)

Negative control : CTLL-2 and Jurkat (PMID : 19147785).

The molecular weight observed is consistent with what has been described in the literature (PMID : 26516065; PMID : 14530327)

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

All lanes:

Western blot - Anti-CD19 antibody [RM1166] (ab317335) at 1/1000 dilution

Lane 1:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

Daudi (human Burkitts lymphoma lymphoblast) whole cell lysate at 20 µg

Lane 3:

Ramos (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 4:

NAMALWA (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 5:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 6:

A20 (mouse reticum sarcoma B lymphocyte) whole cell lysate at 20 µg

Lane 7:

WEHI-231 (mouse B cell lymphoma B lymphocyte ) whole cell lysate at 20 µg

Lane 8:

CTLL-2 (mouse T lymphocyte) whole cell lysate at 20 µg

Lane 9:

Human tonsil tissue lysate at 20 µg

Lane 10:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 60-120 kDa,36 kDa

false

Exposure time: 26s