重组Anti-CD30抗体[EPR4102]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD30 antibody [EPR4102] (AB134080)

Immunohistochemical analysis of paraffin embedded Human Hodgkin's lymphoma tissue labelling CD30 with ab134080 (unpurified) antibody at a dilution of 1/1000. Heat mediated antigen retrieval was perfomed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD30 antibody [EPR4102] (AB134080)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue sections labeling CD30 with purified ab134080 at 1/2000 dilution (0.82 μg/mL). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Lab

Western blot - Anti-CD30 antibody [EPR4102] (AB134080)

The 90 KD band is precursor and 120 KD is the mature transmembrane form as is described in PMID : 11120787.
Raji and Daudi are negative for CD30 as is described in PMID : 29316337.
This antibody detects high background.

Blocking buffer and concentration : 5% NFDM/TBST

Diluting buffer and concentration : 2% BSA/TBST

All lanes:

Western blot - Anti-CD30 antibody [EPR4102] (ab134080) at 1/200 dilution

Lane 1:

KARPAS-299 (Human anaplastic large cell lymphoma) whole cell lysates at 20 µg

Lane 2:

HDLM-2 (Human Hodgkin lymphoma) whole cell lysates at 20 µg

Lane 3:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 20 µg

Lane 4:

Daudi (Human Burkitt's lymphoma lymphoblast) whole cell lysates at 20 µg

Secondary

All lanes:

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

Predicted band size: 64 kDa

Observed band size: 120 kDa,90 kDa

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Exposure time: 60s

• WB

CiteAb

Western blot - Anti-CD30 antibody [EPR4102] (AB134080)

CD30 western blot using anti-CD30 antibody [EPR4102] ab134080. Publication image and figure legend from Wang, R., Li, L., et al., 2018, Mol Oncol, PubMed 29316337.

ab134080 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab134080 please see the product overview.

Anti‐CD30‐LDP retains binding and functional properties of parent anti‐CD30 antibody in vitro. (A) Binding activity of anti‐CD30‐LDP and anti‐CD30 antibody to recombinant human CD30 by ELISA. (B) Surface plasmon resonance (SPR) sensorgrams of anti‐CD30‐LDP binding with CD30 antigen. Various concentrations of antigen (human recombinant CD30, 2.03‐65 nm) were applied on CM5 chip with immobilized anti‐CD30‐LDP fusion protein. Each sensorgram represents different concentrations of CD30. The CD30 at 2.03 nm was injected twice to verify reproducibility. (C) Western blot analysis of CD30 expression levels on different cancer cells. (D) Binding activity to different cell lines. The cells were incubated with anti‐CD30‐LDP of 10 μg·mL−1 and FITC‐conjugated anti‐human Fc second antibody, and then, cell‐bound fluorescence was determined by FACS analysis. The horizontal axis represents the values of MFI. (E) Binding curves of increasing concentrations of anti‐CD30‐LDP (0.001‐3 μg·mL−1) to different cancer cells by FACS analysis. The vertical axis represents the MFI values. (F) In vitro ADCC analysis of anti‐CD30‐LDP and anti‐CD30 antibody. The ADCC assay was performed using PBMCs as effector cells and Karpas299 or L540 cells as target cells at a ratio of 40  : 1. The concentration of anti‐CD30‐LDP was 10 μg·mL−1. Meanwhile, the cetuximab and the target cell line A431 were used as a positive control.

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