Recombinant Anti-Granzyme A antibody [EPR20161] (ab209205)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR20161] to Granzyme A
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt
Reacts with: Human

Product name

Anti-Granzyme A antibody [EPR20161]
See all Granzyme A primary antibodies
Description

Rabbit monoclonal [EPR20161] to Granzyme A
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment within Human Granzyme A aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P12544
Positive control
- WB: NK-92 whole cell lysate; Human spleen, tonsil, lymph node and lung lysates. IHC-P: Human NK/T lymphoma, spleen and endometrium tissues. ICC/IF: NK-92 cells. Flow Cyt: NK-92 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20161
Isotype

IgG
Research areas

Alternative Versions
- Anti-Granzyme A antibody [EPR20161] - BSA and Azide free (ab251499)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)
Recombinant Protein
- Recombinant human Granzyme A protein (ab157288)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Our Abpromise guarantee covers the use of ab209205 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/1000. Detects a band of approximately 28 kDa (predicted molecular weight: 29 kDa).
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/100.
Flow Cyt		1/70.

Function

This enzyme is necessary for target cell lysis in cell-mediated immune responses. It cleaves after Lys or Arg. Cleaves APEX1 after 'Lys-31' and destroys its oxidative repair activity. Involved in apoptosis.
Sequence similarities

Belongs to the peptidase S1 family. Granzyme subfamily.
Contains 1 peptidase S1 domain.
Cellular localization

Secreted. Cytoplasmic granule.
Target information above from: UniProt accession P12544 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3001 Human
- Omim: 140050 Human
- SwissProt: P12544 Human
- Unigene: 90708 Human
Alternative names
- CTL tryptase antibody
- CTLA3 antibody
- Cytolytic t cell and natural killer cell specific trypsin like serine protease antibody
- Cytotoxic T lymphocyte associated serine esterase 3 antibody
- Cytotoxic T lymphocyte proteinase 1 antibody
- Cytotoxic T-lymphocyte proteinase 1 antibody
- Fragmentin 1 antibody
- Fragmentin-1 antibody
- GRAA_HUMAN antibody
- Granzyme 1 cytotoxic T lymphocyte associated serine esterase 3 antibody
- Granzyme A (Cytotoxic T lymphocyte associated serine esterase 3; Hanukah factor serine protease) antibody
- Granzyme A (granzyme 1, cytotoxic T lymphocyte associated serine esterase 3) antibody
- Granzyme A antibody
- Granzyme A precursor antibody
- Granzyme-1 antibody
- GZMA antibody
- H factor antibody
- Hanukah factor serine protease antibody
- Hanukkah factor antibody
- HF antibody
- HFSP antibody
- TSP1 antibody
see all

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

Immunohistochemical analysis of paraffin-embedded human NK/T lymphoma tissue labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on human NK/T lymphoma is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Granzyme A antibody [EPR20161] (ab209205)

All lanes : Anti-Granzyme A antibody [EPR20161] (ab209205) at 1/10000 dilution

Lane 1 : NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) whole cell lysate
Lane 2 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 29 kDa
Observed band size: 28 kDa
why is the actual band size different from the predicted?

Exposure time: 1 second

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression profile is consistent with what has been described in the literature (PMID:15911377).
Immunocytochemistry/ Immunofluorescence - Anti-Granzyme A antibody [EPR20161] (ab209205)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) cells labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining on NK-92 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Flow Cytometry - Anti-Granzyme A antibody [EPR20161] (ab209205)

Flow cytometric analysis of 4% paraformaldehyde-fixed NK-92 (Human peripheral blood malignant non-Hodgkin's lymphoma cell line) cells labeling Granzyme A with ab209205 at 1/70 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-Granzyme A antibody [EPR20161] (ab209205)

All lanes : Anti-Granzyme A antibody [EPR20161] (ab209205) at 1/1000 dilution

Lane 1 : Human spleen lysate at 10 µg
Lane 2 : Human tonsil lysate at 10 µg
Lane 3 : Human lymph node lysate at 10 µg
Lane 4 : Human lung lysate at 20 µg

Secondary
Lanes 1-2 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 29 kDa
Observed band size: 28 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1: 15 seconds, Lane 2: 3 minutes, Lane 3: 10 seconds, Lane 4: 5 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on human spleen is observed [PMID: 9637500].

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Granzyme A antibody [EPR20161] (ab209205)

Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling Granzyme A with ab209205 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Cytoplasmic staining on some stromal cells of human endometrium is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Granzyme A antibody [EPR20161] (ab209205)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet

Publishing research using ab209205? Please let us know so that we can cite the reference in this datasheet.

ab209205 has been referenced in 1 publication.

Wakiyama H et al. Cytolytic Activity (CYT) Score Is a Prognostic Biomarker Reflecting Host Immune Status in Hepatocellular Carcinoma (HCC). Anticancer Res 38:6631-6638 (2018). PubMed: 30504371

Customer reviews and Q&As

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Recombinant Anti-Granzyme A antibody [EPR20161] (ab209205)

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

Applications

Target

Function

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (1)

Customer reviews and Q&As