Product datasheet

Anti-CD20 antibody [SP32] ab64088
リコンビナント

12 Abreviews 24 References 画像数 16
製品の概要
製品名

Anti-CD20 antibody [SP32]

製品の詳細

Rabbit monoclonal [SP32] to CD20

由来種

Rabbitアプリケーション

適用あり : Flow Cyt (Intra), WB, IP, mIHC, IHC-P, ICC/IF

種交差性

交差種 : Mouse, Rat, Human
交差が予測される動物種 : Cow, Dog

免疫原Synthetic peptide within Human CD20 aa 250 to the C-terminus. The exact sequence isproprietary.
Database link: P11836

ポジティブ･コントロール

IHC-P: Human tonsil tissue. Mouse and rat spleen tissues. ICC: Ramos cells. Flow Cyt (Intra):
WEHI-231 and Ramos cells. WB: Ramos cell lysate.

特記事項

This product is FOR RESEARCH USE ONLY. For commercial use, please contact
partnerships@abcam.com.

製品の特性
製品の状態

Liquid

保存方法

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

バッファー

pH: 7.60
Preservative: 0.1% Sodium azide
Constituents: PBS, 1% BSA

精製度

Protein A/G purified

特記事項（精製）

Purified from TCS by protein A/G.

ポリ /モノ

モノクローナル

クローン名

SP32

アイソタイプ

IgG

アプリケーション1

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab64088の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション特記事項

Abreviews

Flow Cyt (Intra)

1/40.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an
isotype control with this antibody.

WB

(1)

Use at an assay dependent concentration.

IP

1/30.

mIHC

Use at an assay dependent concentration.

IHC-P

(6)

1/100. Perform heat mediated antigen retrieval with citrate buffer
pH 6 before commencing with IHC staining protocol.

ICC/IF(1)

1/100.

ターゲット情報機能

This protein may be involved in the regulation of B-cell activation and proliferation.

組織特異性

Expressed on B-cells.

関連疾患

Defects in MS4A1 are the cause of immunodeficiency common variable type 5 (CVID5)[MIM:613495]; also called antibody deficiency due to CD20 defect. CVID5 is a primary
immunodeficiency characterized by antibody deficiency, hypogammaglobulinemia, recurrent
bacterial infections and an inability to mount an antibody response to antigen. The defect resultsfrom a failure of B-cell differentiation and impaired secretion of immunoglobulins; the numbers ofcirculating B cells is usually in the normal range, but can be low.

配列類似性

Belongs to the MS4A family.

翻訳後修飾

Phosphorylated. Might be functionally regulated by protein kinase(s).

細胞内局在Membrane.

画像2ab64088 at 1/30 dilution immunoprecipitating CD20 in Raji (humanBurkitt's lymphoma B lymphocyte) whole cell lysate. Western blot
was performed from the immunoprecipitate using ab64088 at
1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366), was used for detection at 1/5000 dilution.
Lane 1: Raji (human Burkitt's lymphoma B lymphocyte) whole celllysate 10 μg
Lane 2: ab64088 IP in Raji (human Burkitt's lymphoma Blymphocyte) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab64088 inRaji whole cell lysate
Immunoprecipitation - Anti-CD20 antibody [SP32](ab64088)

Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 8 seconds.

Immunohistochemistry analysis (Formalin/PFA-fixed paraffinembedded sections) of paraformaldehyde fixed mouse spleen
tissue. Stained with ab64088 at 1/100 dilution. Secondary antibody
used was Dako EnVision+ System- HRP Labelled Polymer Anti-R.
Blocking was done with 5% serum for 1 hour at 21°C. The sample
was incubated with the primary antibody and 5% Normal Goat
Serum for 19 hours at 4°C. Antigen retrieval method was heat
mediated Citrate pH 6.0.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-CD20 antibody [SP32](ab64088)
This image is courtesy of an anonymous Abreview

3Immunohistochemistry analysis (Formalin/PFA-fixed paraffinembedded sections) of paraformaldehyde fixed human palatine
tonsil tissue. Stained with ab64088 at 1/100 dilution. Secondaryantibody used was Dako EnVision+ System- HRP Labelled
Polymer Anti-R. Blocking was done with 5% serum for 1 hour at
21°C. The sample was incubated with the primary antibody and 5%
Normal Goat Serum for 19 hours at 4°C. Antigen retrieval method
was heat mediated Citrate pH 6.0.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-CD20 antibody [SP32](ab64088)
This image is courtesy of an anonymous Abreview

All lanes : Anti-CD20 antibody [SP32] (ab64088) at 1/1000
dilution

Lane 1 : Wild-type Raji cell lysate
Lane 2 : MS4A1 knockout Raji cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Western blot - Anti-CD20 antibody [SP32] (ab64088)

Observed band size: 33 kDa

Western blot: Anti-MS4A1 antibody [SP32] (ab64088) staining at
1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A](ab7291) loading control staining at 1/20000 dilution, shown inmagenta. In Western blot, ab64088 was shown to bind specificallyto MS4A1. A band was observed at 33 kDa in wild-type Raji celllysates with no signal observed at this size in MS4A1 knockout cellline. To generate this image, wild-type and MS4A1 knockout Rajicell lysates were analysed. First, samples were run on an SDS4

PAGE gel then transferred onto a nitrocellulose membrane.
Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20(TBS-T) before incubation with primary antibodies overnight at 4 °C.
Blots were washed four times in TBS-T, incubated with secondary
antibodies for 1 h at room temperature, washed again four times
then imaged. Secondary antibodies used were Goat anti-Rabbit
IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000
dilution.

This image was generated using ab236434, the same clone, butwith a different buffer formulation.
Multiplex immunohistochemistry analysis of formalin/PFA-fixedparaffin-embedded Rat spleen tissue labeling
Sialoadhesin/CD169, CD3 and CD20 with ab312840 at 1/100
dilution, ab16669 at 1:150 dilution and ab236434 at 1:5000
dilution.
Panel A: merged staining of anti-Sialoadhesin/CD169 (red;Opal™690), anti-CD3 (green; Opal™520) and anti-CD20 (gray;Opal™570) on rat spleen.
Panel B: anti-Sialoadhesin/CD169 stained on macrophages.
Multiplex immunohistochemistry - Anti-CD20
antibody [SP32] (ab64088)

Panel C: anti-CD3 stained on T cells.
Panel D: anti-CD20 stained on B cells.
The section was incubated in three rounds of staining: in the orderof ab312840, ab16669, and ab236434 for 30 mins at room
temperature. Each round was followed by a separate fluorescenttyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems
BOND® RX instrument with an Opal™ 4-color kit. Image acquisition
was performed with Leica SP8 confocal microscope.

5This image was generated using ab236434, the same clone, butwith a different buffer formulation.
Multiplex immunohistochemistry analysis of formalin/PFA-fixedparaffin-embedded Mouse spleen tissue labeling
Sialoadhesin/CD169, CD3 and CD20 with ab312840 at 1/100
dilution, ab16669 at 1:150 dilution and ab236434 at 1:5000
dilution.
Panel A: merged staining of anti-Sialoadhesin/CD169 (red;Opal™690), anti-CD3 (green; Opal™520) and anti-CD20 (gray;Opal™570) on rat spleen.
Panel B: anti-Sialoadhesin/CD169 stained on macrophages.
Multiplex immunohistochemistry - Anti-CD20
antibody [SP32] (ab64088)

Panel C: anti-CD3 stained on T cells.
Panel D: anti-CD20 stained on B cells.
The section was incubated in three rounds of staining: in the orderof ab312840, ab16669, and ab236434 for 30 mins at room
temperature. Each round was followed by a separate fluorescenttyramide signal amplification system.
The immunostaining was performed on a Leica Biosystems
BOND® RX instrument with an Opal™ 4-color kit. Image acquisition
was performed with Leica SP8 confocal microscope.

Flow cytometry analysis of Ramos (human Burkitt's lymphoma)labeling CD20 with purified ab64088 at 1/40 dilution (7.78 µg/ml)(red). Cells were fixed with 4% paraformaldehyde andpermeabilised with 90% methanol. Goat anti rabbit IgG (AlexaFluor® 488, ab150077) at 1/2000 dilution was used as a secondary
antibody. Isotype control -Rabbit monoclonal IgG (ab172730)(Black). Unlableled control -Unlabelled cells (blue).

Flow Cytometry (Intracellular) - Anti-CD20 antibody[SP32] (ab64088)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of Rat spleen tissue sections labeling CD20 withab64088 at 1/100 dilution (3.11 µg/ml). Heat mediated antigen
retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for
10mins. Rabbit specific IHC polymer detection kit HRP/DAB(ab209101) was used as the secondary antibody. Hematoxylin wasused as a counterstain. Positive staining on rat spleen, performed
on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64088 for 30 mins at room
temperature

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-CD20 antibody [SP32](ab64088)

Immunocytochemistry/ Immunofluorescence analysis of Ramos(human Burkitt's lymphoma B lymphocyte) cells labeling CD20 withpurified ab64088 at 1/100 (3.1 µg/ml). Cells were fixed in 4%
paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells
were counterstained with Ab195889 Anti-alpha Tubulin antibody[DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/ml).
Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as thesecondary antibody at 1/1000 (2 µg/ml) dilution. DAPI was used asnuclear counterstain. PBS instead of the primary antibody was usedas the secondary antibody only control.

Immunocytochemistry/ Immunofluorescence - AntiCD20 antibody [SP32] (ab64088)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of Mouse spleen tissue sections labeling CD20
with ab64088 at 1/100 dilution (3.11 µg/ml). Heat mediated antigenretrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for
10mins. Rabbit specific IHC polymer detection kit HRP/DAB(ab209101) was used as the secondary antibody. Hematoxylin wasused as a counterstain. Positive staining on mouse spleen,
performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64088 for 30 mins at room
temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-CD20 antibody [SP32](ab64088)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of Human tonsil tissue sections labeling CD20with ab64088 at 1/100 dilution (3.11 µg/ml). Heat mediated antigen
retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for
10mins. Rabbit specific IHC polymer detection kit HRP/DAB(ab209101) was used as the secondary antibody. Hematoxylin wasused as a counterstain. Positive staining on human tonsil,
performed on a Leica Biosystems BOND™ RX instrument.
The section was incubated with ab64088 for 30 mins at room
temperature.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-CD20 antibody [SP32](ab64088)

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Immunohistochemical analysis of CD20 expression in formalin
fixed, paraffin embedded human tonsil tissue using ab64088 at a1/100 dilution.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-CD20 antibody [SP32](ab64088)

Flow cytometry analysis of WEHI-231 (mouse lymphoblast B celllymphoma) labeling CD20 with purified ab64088 at 1/40 dilution(7.78 µg/ml) (red). Cells were fixed with 4% paraformaldehyde andpermeabilised with 90% methanol. Goat anti rabbit IgG (AlexaFluor® 488, ab150077) at 1/2000 dilution was used as a secondaryantibody. Isotype control -Rabbit monoclonal IgG (ab172730)(Black). Unlableled control -Unlabelled cells (blue).

Flow Cytometry (Intracellular) - Anti-CD20 antibody[SP32] (ab64088)

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Anti-CD20 antibody [SP32] (ab64088) at 1/100 dilution + lysate
prepared from Ramos cells

Observed band size: 33 kDa

Western blot - Anti-CD20 antibody [SP32] (ab64088)

Flow cytometric analysis of rabbit anti-CD20 (SP32) antibody
ab64088(1/100) in Jurkat cells (green) compared to negative
control of rabbit IgG (blue).

Flow Cytometry (Intracellular) - Anti-CD20 antibody[SP32] (ab64088)

Anti-CD20 antibody [SP32] (ab64088)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"

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