Goat anti-rabbit IgG H&L (Alexa Fluor® 647) is a preadsorbed secondary antibody with a maximum absorption wavelength of 650nm and a maximum emission wavelength of 665nm. Ideal for fluorescent cell and tissue imaging. Suitable for ICC/IF, IHC, flow cytometry and ELISA.
- Preadsorbed to prevent cross-reactivity and reduce background staining
- Intense fluorescence and high photostability allowing more time for image capture especially when detecting low abundance targets
- Minimal spectral overlap with other Alexa Fluor® dyes makes this ideal for use in multi-color analysis
- Cited in over 170 publications
Images
Publications
Filter by
- Renal failure 46:23007272024CXC chemokine receptor 7 ameliorates renal fibrosis by inhibiting β-catenin signaling and epithelial-to-mesenchymal transition in tubular epithelial cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPing Meng,Chunli Liu,Jingchun Li,Ping Fang,Bo Yang,Wei Sun,Yunfang ZhangPubMed 38189094
- Bio-protocol 13:e49082023Quantifying Cell Proliferation Through Immunofluorescence on Whole-Mount and Cryosectioned Regenerating Caudal Fins in African Killifish.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAugusto Ortega Granillo,Robert R Schnittker,Wei Wang,Alejandro Sánchez AlvaradoPubMed 38156030
- Journal of orthopaedic surgery and research 18:9672023Sodium salicylate ameliorates exercise-induced muscle damage in mice by inhibiting NF-kB signaling.Applications:Unspecified applicationReactive species:Unspecified reactive speciesYiming Wang,Yuning Sun,Chunhui Yang,Bing Han,Sining WangPubMed 38098039
- International journal of molecular sciences 24:2023Phosphate (Pi) Transporter PIT1 Induces Pi Starvation in -Containing Vacuole in HeLa Cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWen Yang,Yingxing Feng,Jun Yan,Chenbo Kang,Ting Yao,Hongmin Sun,Zhihui ChengPubMed 38139044
- Life (Basel, Switzerland) 13:2023Mathematical Modeling of Non-Small-Cell Lung Cancer Biology through the Experimental Data on Cell Composition and Growth of Patient-Derived Organoids.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRushan Sulimanov,Konstantin Koshelev,Vladimir Makarov,Alexandre Mezentsev,Mikhail Durymanov,Lilian Ismail,Komal Zahid,Yegor Rumyantsev,Ilya LaskovPubMed 38004368
- Journal of materials science. Materials in medicine 34:542023Unusual effects of a nanoporous gold substrate on cell adhesion and differentiation because of independent multi-branch signaling of focal adhesions.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPeizheng Wu,Kazuya Yanagi,Kazuki Yokota,Masataka Hakamada,Mamoru MabuchiPubMed 37884819
- International journal of ophthalmology 16:1582-15882023Effect of miR-27b-3p and Nrf2 in human retinal pigment epithelial cell induced by high-glucose.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQiao-Ling Lai,Ting Xie,Wei-Dong Zheng,Yan HuangPubMed 37854382
- International journal of molecular medicine 52:2023Arctiin attenuates iron overload‑induced osteoporosis by regulating the PI3K/Akt pathway.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMiao Li,Zhaofeng Pan,Qi He,Jiacong Xiao,Baihao Chen,Fanchen Wang,Pan Kang,Haoran Luo,Jianliang Li,Jiaxu Zeng,Shaocong Li,Junzheng Yang,Haibin Wang,Chi ZhouPubMed 37800616
- PloS one 18:e02923172023LucFlow: A method to measure Luciferase reporter expression in single cells.Applications:Unspecified applicationReactive species:Unspecified reactive speciesSunil Nooti,Madison Naylor,Trevor Long,Brayden Groll, ManuPubMed 37792708
- Stem cell research & therapy 14:2632023Human umbilical cord-derived mesenchymal stem cells ameliorate perioperative neurocognitive disorder by inhibiting inflammatory responses and activating BDNF/TrkB/CREB signaling pathway in aged mice.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPenghui Wei,Min Jia,Xiangyi Kong,Wenyuan Lyu,Hao Feng,Xinyi Sun,Jianjun Li,Jian-Jun YangPubMed 37735415
Key facts
- Host species
- Goat
- Target species
- Rabbit
- Target isotype
- IgG
- Target specificity
- Heavy & Light chains
- Minimal cross-reactivity
- Mouse, Rat, Human
- Conjugation
- Alexa Fluor® 647
- Excitation/Emission
- Ex: 650nm, Em: 665nm
- Applications
- ELISA, IHC-Fr, IHC-P, Flow Cyt, ICC/IF
- Clonality
- Polyclonal
Abcam Recommends
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISA | Reactivity Reacts | Dilution info - | Notes - |
Application IHC-Fr | Reactivity Reacts | Dilution info - | Notes - |
Application IHC-P | Reactivity Reacts | Dilution info - | Notes - |
Application Flow Cyt | Reactivity Reacts | Dilution info 1/2000 | Notes ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control to complement this secondary antibody. |
Application ICC/IF | Reactivity Reacts | Dilution info 1/200.00000 - 1/1000.00000 | Notes - |
Associated Products
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Target data
Alternative names
Ig gamma 1 chain C region, Ig gamma 3 chain C region, Ig gamma 4 chain C region, Ig gamma chain C region, Ig gamma-2 chain C region, Ig kappa-b4 chain C region, Ig kappa-b5 chain C region, Ig kappa-b9 chain C region, Ig lambda chain C region, Immunoglobin heavy constant gamma 1, Immunoglobulin G, K-BAS
Recommended products
Goat anti-rabbit IgG H&L (Alexa Fluor® 647) is a preadsorbed secondary antibody with a maximum absorption wavelength of 650nm and a maximum emission wavelength of 665nm. Ideal for fluorescent cell and tissue imaging. Suitable for ICC/IF, IHC, flow cytometry and ELISA.
- Preadsorbed to prevent cross-reactivity and reduce background staining
- Intense fluorescence and high photostability allowing more time for image capture especially when detecting low abundance targets
- Minimal spectral overlap with other Alexa Fluor® dyes makes this ideal for use in multi-color analysis
- Cited in over 170 publications
Key facts
- Description
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed
- Host species
- Goat
- Target species
- Rabbit
- Target isotype
- IgG
- Target specificity
- Heavy & Light chains
- Minimal cross-reactivity
- Mouse, Rat, Human
- Conjugation
- Alexa Fluor® 647
- Excitation/Emission
- Ex: 650nm, Em: 665nm
- Applications
- ELISA, IHC-Fr, IHC-P, Flow Cyt, ICC/IF
- Clonality
- Polyclonal
- Pre-adsorbed
- Yes
- Specificity
By immunoelectrophoresis and ELISA this antibody reacts specifically with rabbit IgG and with light chains common to other rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins. Reduced cross-reactivity to bovine, chicken, horse, human, mouse, pig, and rat IgG was detected. This antibody may cross react with IgG from other species.
- Isotype
- IgG
- Concentration
Properties
- Form
- Liquid
- Storage buffer
Preservative: 0.02% Sodium azide
Constituents: PBS, 23% Glycerol (glycerin, glycerine), 1% BSA- Purification technique
- Affinity purification Immunogen
- Purification notes
Antiserum was cross adsorbed using a human, mouse and rat immunosorbents to remove cross reactive antibodies. The antibody to rabbit IgG was isolated by affinity chromatography using antigen coupled to agarose beads.
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage duration
- 1-2 weeks
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- -20°C
- Aliquoting information
- Upon delivery aliquot
- Storage information
- Avoid freeze / thaw cycle, Stable for 12 months at -20°C, Store in the dark
Notes
Fluorochrome chart- a complete quick and easy guide to help you select the most appropriate fluorochromes for your next experiment.
When it comes to advancing your immunohistochemistry (IHC) research, Abcam offers a comprehensive suite of IHC kits and secondary antibodies, designed to deliver precise and reliable results.
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
41 product images
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
ICC/IF image of Anti-beta Tubulin antibody - Loading Control ab6046 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (Anti-beta Tubulin antibody - Loading Control ab6046, 1µg/ml) overnight at +4°C. The secondary antibody (red) was ab150083 Alexa Fluor® 647 goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
HeLa cells showing negative staining by ICC/IF using only secondary antibody. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The secondary antibody (red) was ab150083 Alexa Fluor® 647 goat anti-rabbit IgG (H+L) used at 2µg/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
This image is courtesy of Dr. Shaohua LiImmunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Image: Courtesy of Dr. Shaohua Li, UMDNJ-Robert Wood Johnson Medical School
Sample: mouse embryonic stem cell-differentiated embryoid bodies (EBs)
Preparation:
Fix in 3%PFA in PBS for 30 min at RTIncubate in 7.5% sucrose-PBS for 3h at RTIncubate in 15% sucrose-PBS at 4 degree Celsius overnightEmbed the EBs in tissue-Tek OCT compoundCut frozen sections to 4-20 μm thickness
Primary antibody 1: Mouse anti-Ki67, 1:50
Primary antibody 2: Rabbit anti-laminin, 1:400
Secondary antibody 1: Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594) pre-adsorbed (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120), 1:200Secondary antibody 2: Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 647) pre-adsorbed (ab150083), 1:200
Nuclei were counterstained with Syto 16
Alexa Fluor® - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of A375 (human malignant melanoma epithelial cell) (Right) / Jurkat (human T cell leukemia T lymphocyte from peripheral blood) (Left) cells labelling OSMR with Anti-OSMR antibody [RM2085] ab324099 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: Jurkat.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Human purified neutrophils cells labelling IL-5RA with Anti-IL-5RA antibody [EPR28313-257] ab324011 at 1/500 dilution (0.1 ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Isotype (Left) / 293T cells transfected with a human SIGLEC6 expression vector containing a GFP-tag® (Middle) / 293T cells transfected with an empty expression vector containing a GFP-tag® (Right) cells labelling SIGLEC6 with Anti-SIGLEC6 antibody [RM2081] ab323795 at 1/5000 dilution (0.01 ug).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of A-172 (human brain glioblastoma cell, Right) / Daudi (human Burkitt's lymphoma lymphoblast, Left) cells labelling IL-13 receptor alpha 2 with Anti-IL-13 receptor alpha 2 antibody [EPR29921-581] ab324048 at 1/500 dilution (0.1 ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Negative control: Daudi.
Gated on viable cells.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A549 (human lung carcinoma epithelial cell) treated with 10ng/mL IFN-alpha 1 for 16 hours (Green) / Untreated A549 (Magenta) cells labelling IFITM2+IFITM3 with Anti-IFITM2+IFITM3 antibody [EPR28942-88] ab323747 at 1/50 dilution (1 ug) / Magenta and Green compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) and Grey isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (upper left) / 293T (human embryonic kidney epithelial cell) cells transfected with an empty expression vector containing a Myc tag (upper middle) / 293T cells transfected with a IFITM1 expression vector containing a GFP tag (down left) / 293T cells transfected with a IFITM2 expression vector containing a GFP tag (down middle) / 293T cells transfected with a IFITM3 expression vector containing a GFP tag(down right) cells labelling IFITM2+IFITM3 with Anti-IFITM2+IFITM3 antibody [EPR28942-88] ab323747 at 1/5000 dilution (0.01 ug) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of MEF (mouse embryo fibroblast, Left) / A20 (mouse reticulum sarcoma B lymphocyte, Right) cells labelling TIM 1 with Anti-TIM 1 antibody [RM1294] ab323414 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Low expression: MEF.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Mouse bone marrow cells labelling CD64 with Anti-CD64 antibody [EPR26480-569] ab323507 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Cells were co-stained with anti mouse CD11b conjugated to Brilliant Violet 421.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Mouse bone marrow cells labelling CD64 with Anti-CD64 antibody [EPR26480-569] ab323507 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Cells were co-stained with anti mouse CD3 conjugated to Alexa Fluor®488.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized A431 (human epidermoid carcinoma epithelial cell) labelling Cytokeratin 1 with Anti-Cytokeratin 1 antibody [EPR17744] ab185628 at 1/500 (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Mouse bone marrow cells labelling Leptin Receptor with Anti-Leptin Receptor antibody [EPR28293-79] ab318272 at 1/200 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Mouse bone marrow are co-stained with CD45-CD31-TER119 conjugated FITC (530/30BP).
Gated on viable cell.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of K-562 (human lung carcinoma epithelial cell) (Right) / Ramos (human Burkitt's lymphoma B lymphocyte) (Left) cells labeling EMR2 with ab323321 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: Ramos (PMID: 11994511)
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling TIGIT with Anti-TIGIT antibody [EPR28146-94] ab321793 at 1/500 dilution (0.1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD45RO conjugated to Pacific Blue.
Gated on viable CD3+ T Lymphocytes.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Left) / 293T cells transfected with a human HA Tag expression vector containing a GFP tag (Middle) / 293T cells transfected with an empty expression vector containing a GFP tag (Right) cells labelling HA tag with Anti-HA tag antibody [RM1058] ab314237 at 1/5000 dilution (0.01 ug)/Middle and Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control.
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of NCI-H460(human Large Cell Lung Cancer epithelial cell, Left) / Raji(human Burkitt's lymphoma B lymphocyte, Right) cells labelling IL-4R with Anti-IL-4R antibody [RM2076] ab323370 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control.
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cell.
Negative control: NCI-H460.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of MCF7(human breast adenocarcinoma epithelial cell, Left) / Daudi(human Burkitt's lymphoma lymphoblast, Right) cells labelling IL-4R with Anti-IL-4R antibody [RM2076] ab323370 at 1/5000 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cell.
Negative control: MCF7.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of MEF (mouse embryo fibroblast, Left) / A20 (mouse reticulum sarcoma B lymphocyte, Right) cells labelling TIM 1 with Anti-TIM 1 antibody [EPR28341-72] ab316854 at 1/500 dilution (1µg) (Red) followed by a Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) secondary antibody used at a 1/5000 dilution compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) ( Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Low expression: MEF.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of T-47D (human ductal breast epithelial tumor epithelial cell) (Right) / A549 (human lung carcinoma epithelial cell) (Left) cells labelling Nectin-4 with Anti-Nectin-4 antibody [RM1257] ab322927 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Low expression: A549 (PMID:21901103).
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of DLD-1 (human colorectal adenocarcinoma epithelial cell) (Right) / HeLa (human cervical adenocarcinoma epithelial cell) (Left) cells labelling Nectin-4 with Anti-Nectin-4 antibody [RM1257] ab322927 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Low expression: HeLa (PMID:21901103).
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of MCF7 (human breast adenocarcinoma epithelial cell) (Right) / T-47D (human ductal breast epithelial tumor epithelial cell) (Left) cells labelling GPCR RDC1/CXCR-7 with Anti-GPCR RDC1/CXCR-7 antibody [EPR29064-22] ab322663 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: T-47D (PMID: 25168820).
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of LNCap (human prostate carcinoma epithelial cell) (Right) / HT-29 (human colorectal adenocarcinoma epithelial cell) (Left) cells labelling GPCR RDC1/CXCR-7 with Anti-GPCR RDC1/CXCR-7 antibody [EPR29064-22] ab322663 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: HT-29 (PMID: 20388803).
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with Anti-Granzyme A antibody [EPR29034-79] ab321992 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD14 conjugated to Pacific blue.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) (Right) / 293T (human embryonic kidney epithelial cell) (Left) cells labelling Granzyme A with Anti-Granzyme A antibody [EPR29034-79] ab321992 at 1/500 dilution (0.01ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Negative control: 293T.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with Anti-Granzyme A antibody [EPR29034-79] ab321992 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD56 conjugated to PE.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Human PBMC (human peripheral blood mononuclear cell) cells labelling Granzyme A with Anti-Granzyme A antibody [EPR29034-79] ab321992 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD8a conjugated to Pacific blue.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Isotype (Upper Left) / 293T (human embryonic kidney epithelial cell) transfected with an Granzyme A expression vector containing a myc-His-tag® (Upper Middle) / 293T cells transfected with an empty expression vector containing a myc-His-tag® (Upper Right) / 293T cells transfected with homologous human Granzyme expression vector (GrzmB, GrzmH, GrzmK, GrzmM) containing a myc-His-tag® (Lower). cells labelling Granzyme A with Anti-Granzyme A antibody [EPR29034-79] ab321992 at 1/50 dilution (1ug) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were first stained with rabbit IgG or Anti-Granzyme A antibody [EPR29034-79] ab321992. After fixation and permeability, cells stained with anti-myc tag conjugated to Alexa Fluor® 647.
Crossreactivity with protein Granzyme B, Granzyme H, Granzyme K, Granzyme M were fully tested.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Human PBMC (human peripheral blood mononuclear cell) cells labelling CLEC2D with Anti-CLEC2D antibody [EPR27405-92] ab319140 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti human CD14 conjugated to FITC.
Gated on viable cells.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Daudi (human Burkitt's lymphoma lymphoblast) (Right) / MCF7 (human breast adenocarcinoma epithelial cell) (Left) cells labelling IL-4R with Anti-IL-4R antibody [EPR28312-39] ab320080 at 1/5000 dilution (0.01ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: MCF7 (PMID: 15331327)
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Isotype (Left) / WT Mouse bone marrow (middle) / Prx1 KO Mouse bone marrow (Right) cells labelling Leptin Receptor with Anti-Leptin Receptor antibody [EPR28293-79] ab318272 at 1/200 dilution (0.1ug) / Middle and Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/2000 dilution was used as the secondary antibody.
Mouse bone marrow are co-stained with CD45-CD31-TER119 conjugated FITC (530/30BP).
Gated on viable cell.
The result was kindly provided by Dr Bo Shen's lab, NIBS.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) (Right) / NIH/3T3 (mouse embryonic fibroblast) (Left) cells labelling CSF-1-R with Anti-CSF-1-R antibody [EPR29702-379] ab319160 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
The experiment was conducted on freshly cultured cells and cells were stained at low temperatures (2 - 8°C) on ice to preserve the surface expression of CD115, since CD115 is prone to rapid internalization under ambient room temperature conditions (PMID: 21466808).
Negative control: NIH/3T3 (PMID: 36555673).
Immunocytochemistry/ Immunofluorescence - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling MERS Spike glycoprotein with Anti-MERS Spike glycoprotein antibody [abd144] ab322146 at 1/500 (1.92 ug/ml) dilution, followed by Goat anti-Human IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor® 488) antibody at 1/1000 2 ug/ml dilution (Green).
Confocal image showing cytoplasmic staining in 293T cells transfected with a MERS-CoV S protein expression vector containing a myc-His-tag®(shown in green) and no staining in 293T cells transfected with a SARS-CoV-2 Spike glycoprotein expression vector containing a myc-His-tag®. The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).Anti-Myc tag antibody ab9106 Anti-Myc Rabbit polyclonal antibody was used to counterstain tubulin at 1/200 2.5 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat anti-Human IgG (H+L) Cross-Adsorbed Secondary Antibody (Alexa Fluor® 488) at 1/1000 2 ug/ml dilution.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Mouse bone marrow cells labelling CCR3 with Anti-CCR3 antibody [EPR27419-30] ab318266 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti-CD11b conjugated to Brilliant Violet 421.
Gated on viable cells.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Mouse bone marrow cells labelling CCR3 with Anti-CCR3 antibody [EPR27419-30] ab318266 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Cells were co-stained with anti-CD3 cojugated to Alexa Fluor®488.
Gated on viable cells.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) (Right) /
A20 (mouse reticulum sarcoma B lymphocyte) (Left) cells labelling CCR3 with Anti-CCR3 antibody [EPR27419-30] ab318266 at 1/50 dilution (1ug) / Magenta compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Low expression: A20.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of No-GFP-CD16.NK-92 (CD16 ectopically expressed natural killer(NK-92®) cell with no GFP tag) (right) / PC-3 (human prostate adenocarcinoma epithelial cell) (Left) cells labelling TIGIT with Anti-TIGIT antibody [EPR28146-94] ab321793 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Gated on viable cells.
Negative control: PC-3.
Flow Cytometry (Intracellular) - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized LNCaP (human prostate carcinoma epithelial cell) (Right) / PC-3 (human prostate adenocarcinoma epithelial cell) (Left) cells labelling Prostate Specific Antigen with Anti-Prostate Specific Antigen antibody [RM1268] ab322442 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Low expression: PC-3.
Flow Cytometry - Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150083)
Flow cytometric analysis of Jurkat (human T cell leukemia T lymphocyte from peripheral blood, Left) /A-204 (human muscle rhabdomyosarcoma cell, Right) cells labelling HHLA2 with Anti-HHLA2 antibody [RM2075] ab323612 at 1/5000 dilution (0.01 ug) / Right compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).
Goat anti-Rabbit IgG (Alexa Fluor® 647, ab150083) at 1/5000 dilution was used as the secondary antibody.
Negative control: Jurkat (PMID: 25549724).
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