The primary antibodies used for western blotting and immunofluorescence analysis included: SDCCAG8 (WB 1:2 000, IHC 1:50, 13471-1-AP; Proteintech Group, USA), GRK1 (1:400, 24606-1-AP; Proteintech Group, USA), rhodopsin (1:400, 1D4, clone D4B9B; Cell Signaling Technology, CST, USA), S-opsin (1:300, Ab5407; Abcam, UK), GFP (1:3 000, 50430-2-AP; Proteintech Group, USA), GAPDH (1:5 000, 10494-1-AP; Proteintech Group, USA), β-actin (1:5 000, 20536-1-AP; Proteintech Group, USA), PDE6B (1:400, T13343; Thermo, USA), Alexa Fluor 594 conjugated peanut agglutinin (PNA) (1:200, L32459; Thermo, USA), cone arrestin (1:300, AB15282; Sigma, USA), and anti-alpha tubulin (acetyl K40) (1:1 000, ab24610; Abcam, UK). Secondary antibodies included: goat anti-rabbit Alexa Fluor 488 and 594 (1:1 000; Invitrogen, USA), goat anti-mouse Alexa Fluor 594 (1:1 000; Invitrogen, USA), and HRP-conjugated Affinipure goat anti-rabbit IgG (H+L) (1:5 000, SA00001-2; Proteintech Group, USA).
Ab5407
Manufactured by Abcam
Sourced in United Kingdom
AB5407 is a rabbit monoclonal antibody that recognizes the human Cyclin A protein. Cyclin A is a key regulator of the cell cycle and plays a crucial role in the transition from the G1 to the S phase and from the G2 to the M phase. This antibody can be used for various research applications, such as Western blotting, immunoprecipitation, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
A11008, by Thermo Fisher Scientific (2 mentions)
Dmi6000 fluorescent microscope, by Leica (2 mentions)
Ab5405, by Merck Group (2 mentions)
Mab5356, by Merck Group (2 mentions)
S 2000, by Vector Laboratories (2 mentions)
Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (2 mentions)
Ab290, by Abcam (2 mentions)
Alexa fluor 594, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 and 594, by Thermo Fisher Scientific (1 mentions)
Ab15282, by Merck Group (1 mentions)
Ab24610, by Abcam (1 mentions)
β actin, by Proteintech (1 mentions)
Affinipure goat anti rabbit igg h l, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
Ab32042, by Abcam (1 mentions)
Ab15580, by Abcam (1 mentions)
Ab223075, by Abcam (1 mentions)
Ix71 inverted microscope, by Olympus (1 mentions)
As038, by ABclonal (1 mentions)
6 protocols using ab5407
1
Immunofluorescence and Western Blot Analysis of Retinal Proteins
2
Histological Preparation of Rhabdomys pumilio Retinas
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Rhabdomys pumilio were anaesthetised with urethane and perfused with 4% paraformaldehyde (methanol free). Eyes were then stored in methanol-free 4% paraformaldehyde prior to further processing. For retinal wholemounts, retinas were dissected from fixed eyes and immunohistochemistry performed on free-floating retinas. For retinal sections, whole eyes were embedded in Historesin and were sectioned at 5 µm thickness. For general histology, fixed eyes were dehydrated through a graded series of alcohols and infiltrated with Technovit 7100 (Historesin TAAB Labs UK). Blocks were sectioned at 5 µm and mounted onto clean slides, stained with Cresyl Violet and coverslipped under DPX. For immunohistochemistry, R. pumilio retinas were labelled using polyclonal antibodies against MWS/LWS opsin raised in chicken (PA1-9517, ThermoFisher; 1:250 dilution) and against SWS opsin raised in rabbit (AB5407, Abcam; 1:300 dilution), and left overnight at room temperature. Tissues were then washed and incubated for 2 h in fluorescent secondary antibodies at a dilution of 1:2000 made up of 2% NDS, 3% Triton X-100 and PBS. Sections were imaged with an Axio Imager.D2 upright microscope and captured using a Coolsnap Hq2 camera (Photometrics) through Micromanager software v1.4.23.
3
Retinal Cell Type Quantification
4
Retinal Cell Type Quantification
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Immunohistochemistry analysis was performed on 10 µm paraffin embedded serial sections from the enucleated mouse eyes as described in our previous studies [72 ]. At minimum 100 µm of retina/sample was evaluated by IHC. Briefly, sections were blocked with 2% normal horse serum (#S-2000 VectorLabs, CA) in PBS, and incubated with the following cell type-specific primary antibodies in a 1:200 dilution: rhodopsin (mouse monoclonal, Millipore MAB5356); green/red opsin (rabbit polyclonal, Millipore AB5405); blue opsin (rabbit polyclonal, Millipore AB5407); GFP (1:500, rabbit polyclonal, Abcam ab290). The following day, sections were rinsed with PBS and incubated with the corresponding secondary antibody (1:400 Alexa fluor 488 goat antirabbit, Invitrogen A11008) and nuclei were stained with 4,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI). Over 500 μm of sections/animal were visualized and representative images of IHC labeling were captured using a Leica DMI6000 fluorescent microscope equipped with the appropriate bandpass filter for each fluorochrome. Cell counts were performed in a double-blinded manner over 100 μm retinal area (N = 10/strain/experimental group).
5
Immunohistochemical Analysis of Colon Cancer
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Both normal and colon cancer tissues collected from mice were fixed for 24 h using 4% paraformaldehyde. Then, immunohistochemical staining for SRCIN1, cleaved caspase 3, β-catenin and Ki-67 was performed by incubation with the following primary antibodies: SRCIN1 (ab5407, 1:200; Abcam), cleaved caspase 3 (ab32042, 1:100; Abcam), β-catenin (ab223075, 1:200; Abcam), and Ki-67 (ab15580, 1:200; Abcam). Finally, the IX71 inverted microscope (Olympus) was used to acquire images.
6
SRCIN1 Expression in NSCLC Tissues
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Initially, NSCLC tumors and paracancerous tissues were fixed with 4% paraformaldehyde for 24 h, after which they were subjected to immunohistochemical (IHC) staining for SRCIN1 using primary anti-SRCIN1 (abcam, ab5407; 1 : 200), and secondary antibody HRP Donkey Anti-Rabbit IgG (H+L) (ABclonal, AS038, 1 : 1000). Samples were then imaged via light microscope (Olympus, Tokyo, Japan).
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