Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor 488) Conjugation: Alexa Fluor® 488. Ex: 495nm, Em: 519nm Host species: Goat Isotype: IgG Suitable for: ICC/IF, Flow Cyt, IHC-P,
ELISA, IHC-Fr
存放说明 Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. Store In the Dark.
存储溶液 Preservative: 0.02% Sodium azide
Constituents: 23% Glycerol (glycerin, glycerine), PBS, 1% BSA
纯化说明 This antibody was isolated by affinity chromatography using antigen coupled to agarose beads.
Fluorochrome chart a complete guide:
A quick and easy guide to help you select the most appropriate fluorochromes for your next experiment.
Please seehere.
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.
The Abpromise guarantee
Abpromise™承诺保证使用ab150077于以下的经测试应用
\"应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
ICC/IF image of beta Tubulin staining in HeLa cells. The cells were 100% methanolfixed (5 min), permeabilized with 0.1% Triton X-100for 5 minutes and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to block non-specific protein-protein interactions. The cells were then incubated with the primary antibody (ab6046, 5 g/ml) overnight at +4 C. The secondary antibody (green) was ab150077 Alexa Fluor 488 goat anti-rabbit IgG (H+L) used at 2 g/ml for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 M.
The negative control (inset) is a secondary-only assay to demonstrate low non-specific binding of the secondary antibody.
Immunocytochemistry/ Immunofluorescence analysis of C6(Rat glial tumor glial cell) cells labeling alpha smooth muscle Actin with purified ab32575 at 1/100 dilution (0.71 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 the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Overlay histogram showing MCF7 cells stained with unpurified ab32063 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32063, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in MCF7 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3(Mouse embryonic fibroblast) cells labeling alpha smooth muscle Actin with purified ab32575 at 1/500 dilution (5.2 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 the secondary antibody at 1/1000 dilution (2 µg/ml) dilution. DAPI was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Estrogen Receptor alpha with purified ab32063 at 1/1000. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. The cells were co-stained with ab7291, a mouse anti-tubulin (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary antibody. Nuclei counterstained with DAPI (blue).
Control 1: primary antibody (1/1000) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
Immunocytochemistry/Immunofluorescence analysis of Raji (human Burkitt\'s lymphoma) cells labelling HLA A with purified ab52922 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
Overlay histogram showing Jurkat cells stained with ab16669 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16669, 1/1000 dilution) for 30 min at 22°C. The secondary antibody Goat anti-rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used at 1/4000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with Purified ab108338 at 1/100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunocytochemistry/Immunofluorescence analysis of A431 (human epidermoid carcinoma) cells labeling alpha smooth muscle Actin (green) with purified ab32575 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were counterstained with ab7291, anti-Tubulin (mouse mAb) at 1/1000 followed by ab150120 Alexa Fluor®594 goat anti-mouse secondary (1/1000). Nuclei were counterstained with DAPI (blue).
For negative control 1, rabbit primary antibody and anti-mouse secondary antibody (ab150120) were used. For negative control 2, ab7291 (mouse primary antibody) was used followed by anti-rabbit secondary antibody (ab150077).
Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling IL-1RA with purified ab124962 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
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 (ab7291, 1 g/ml) and (ab16048, 1 g/ml) overnight at +4 C. The secondary antibodies were ab150115 Alexa Fluor 647 (red) goat anti-mouse IgG (H+L) used at 2 g/ml for 1h and ab150077 Alexa Fluor 488 (green) goat anti-rabbit IgG (H+L) used at 2 g/ml for 1h.DAPI was used to stain the cell nuclei.
Overlay histogram showing HeLa cells stained with ab32356 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32356,1/100 dilution) for 30 min at 22 C. The secondary antibody used was Alexa Fluor 488 goat anti-rabbit IgG (H L) at 1/2000 dilution for 30 min at 22 C. Isotype control antibody (black line) wasrabbit IgG (monoclonal) (1 g/1x10^6 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of 5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
Cross-reactivity of the polyclonal secondary antibody ab182016 was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards at 1 g/ml(50 l/well) and incubatedovernight at 4 C, followed by a 5% BSA blocking step for 2h at RT. ab182016 was then added starting at 1 g/ml and gradually diluted 1/4 (50 l/well), followed by incubation for 2h. For the detection Donkey anti-Goat IgG H L (HRP)(ab6885) was used at 1/10,000 dilution (50 l/well), followed by incubationfor 1h at RT.
For the batch tested, ab182016 showed a cross-reactivity of 5-7% towards Human IgG and below 2% towards Mouse IgG, RatIgG and Chicken IgY.
This data was developed using the unconjugated antibody (ab182016).
Cross-reactivity of Goat anti-Rabbit IgG H L (ab182016)and Goat anti-Rabbit IgG H Lobtained from two different vendors was tested using a sandwich ELISA approach. The wells were coated with the indicated IgG standards (Rabbit, Human, Mouse and Rat) at 1 g/ml (50 l/well) and incubatedovernight at 4 C, followed by a 5% BSA blocking step for 2h at RT. Secondary antibodies were then added starting at 1 g/ml and gradually diluted 1/4 (50 l/well), followed by incubationfor 2h. For the detection Donkey anti-Goat IgG H L (HRP)(ab6885) was used at 1/10,000 dilution (50 l/well), followed by incubationfor 1h atRT. This data is from a representative dilution.
This data was developed using the unconjugated antibody (ab182016).
IHC - Wholemount of Caenorhabditis elegans larvae labelling RNA polymerase II CTD repeat YSPTSPS (phospho S2) with ab5095. The sample was incubated with primary antibody (1/500 in PBS + 3% BSA + 0.1% Triton X-100) for 12 hours at 4 C. ab150077, an Alexa Fluor 488-conjugated goat anti-rabbit IgG polyclonal (1/1000), was used as the secondary antibody.
See Abreview
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Click here to view the general protocols
发表研究结果有使用 ab150077？请让我们知道，以便我们可以引用本数据表中的参考文章。
ab150077 被引用在 1034 文献中.
Xiao M et al. Long non-coding RNA H19 promotes the proliferation, migration and invasion while inhibits apoptosis of hypertrophic scarring fibroblasts by targeting miR-3187-3p/GAB1 axis. Burns 47:654-664 (2021).PubMed: 32888745 Gan L et al. Mesenchymal stem cells promote chemoresistance by activating autophagy in intrahepatic cholangiocarcinoma. Oncol Rep 45:107-118 (2021).PubMed: 33155663 Li Y et al. Songorine promotes cardiac mitochondrial biogenesis via Nrf2 induction during sepsis. Redox Biol 38:101771 (2021).PubMed: 33189984 Wei H et al. miR-34c-5p targets Notch1 and suppresses the metastasis and invasion of cervical cancer. Mol Med Rep 23:N/A (2021).PubMed: 33300051 Niu YT et al. In the presence of TGF-ß1, Asperosaponin VI promotes human mesenchymal stem cell differentiation into nucleus pulposus like- cells. BMC Complement Med Ther 21:32 (2021).PubMed: 33446173 View all Publications for this product
We used this secondary antibody to stain amyloid plaques in human brain tissue sections.
The signals were very clear, specific and strong.
This secondary antibody can be fully recommended.
Conditions:
Anti rabbit-Alexa488, 1:500, 21 °C, 30 min
Sample was a liver slide (FFPE-tissue)
Primary antibody: Rabbit anti-Transferrin
Secondary antibody: Goat anti-rabbit IgG H&L (Alexa Fluor® 488)Abcam ab150077
Blocking buffer: 5% Goat serum
Incubation of secondary antibody: 1:500 at RT for 30 min
Strong fluorescent signal with moderate background noise was observed.
HeLa cells were imaged using ab150077 as a secondary antibody.
Fix cells for 5 min with 4% paraformaldehyde, permeabilize for 5 min with 0.1% Triton-X 100
Primary antibody: ab39688, MYC, 1:100 dilution, RT, 90 min
Secondary antibody: ab150077, 1:500 dilution, RT, 45 min
Used for IF staining of kryo frozen prostate cancer slides.
Primary antibody: TIMP1 (ab211926).
Gives great signal at 1:500 dilution, minimal background.
Inc. time primary and secondary: 30 min at 21°C
This Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) worked well with Anti-GRP78 BiP antibody ab21685 in deer mice skin sections.
Cell: HeLa
Fix & permeabilization: 4% formalin, 0.03% Triton-X100
Blocking: 3% BSA/PBS
1st antibody: anti-Nup133 Rabbit polyclonal antibody (1/500 dil, 3% BSA/PBS)
2nd antibody: Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) (1/2500 dil, 3% BSA/PBS)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
The secondary antibody works well. Based on my experience for IHC-P the dilution factor requires optimization based on the tissue and antigen retrieval method used.
The dilution 1:250 works in my case.
Immunocytochemistry/ Immunofluorescence
Resulted in bright and specific staining.
Cells: PBMCs
Fixation: Acetone
Primary antibody: CD3
Blocking: 5% BSA in PBS
Counterstained with DAPI
Melanoma tissue was stained to see CD3 infiltrates.
Tissue was fixed with acetone for 10 min at -20 °C.
The intensity of the fluorescence was really great.
No Background signal was detected.
I strongly can recommend this antibody.
Application: Immunocytochemistry
Cells were fixed in 2% Paraformaldehyde.
Permeabilization: Triton X-100
Primary antibody: Rabbit polyclonal anti-a-tubulin
Blocking: 5% BSA in PBS
Counterstained with DAPI and actin (555)
Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES
For licensing inquiries, please contact partnerships@abcam.com
