Anti-Granzyme B antibody [EPR8260]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] (AB134933)

Immunohistochemical analysis of paraffin-embedded, formalin-fixed Human Hodgkin's lymphoma tissue, labelling Granzyme B using unpurified ab134933 at a 1/100 dilution. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] (AB134933)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling Granzyme B with Purified ab134933 at 1 : 250 dilution (2.96 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use)was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-Granzyme B antibody [EPR8260] (AB134933)

Flow cytometric analysis of 4% paraformaldehyde fixed No-GFP-CD16.NK-92 Human Natural Killer cells labeling Granzyme B with ab134933 at 1/100 dilution (Red) compared with an Isotype control Rabbit monoclonal IgG (ab172730) (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti rabbit IgG Alexa Fluor® 488 (ab150081), at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme B antibody [EPR8260] (AB134933)

Tissue Microarrays stained for "Anti-Granzyme B antibody [EPR8260]" using "ab134933"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab134933 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-Granzyme B antibody [EPR8260] (AB134933)

Negative control : Jurkat (PMID : 18437383, 25168906).

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Granzyme B antibody [EPR8260] (ab134933) at 1/1000 dilution

Lane 1:

KARPAS-299 (human anaplastic large cell lymphoma), whole cell lysate at 20 µg

Lane 2:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 3:

SR (human pleural effusion lymphoblast, whole cell 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 1/10000 dilution

Predicted band size: 28 kDa

Observed band size: 28 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Granzyme B antibody [EPR8260] (AB134933)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of KARPAS-299 whole cell lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab134933 (1/1000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/100,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique ab134933 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant for temperatures ranging between 4°C, 22°C and 37°C, demonstrating that antibody activity is not affected under these conditions.

All lanes:

Western blot - Anti-Granzyme B antibody [EPR8260] (ab134933) at 1/1000 dilution

All lanes:

KARPAS-299 whole cell lysate at 15 µg with NDFM/TBST

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 1/100000 dilution

false

Exposure time: 20s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-Granzyme B antibody [EPR8260] (AB134933)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.