Cells were fixed with 4% paraformaldehyde (in PBS; Carl Roth) and incubated in blocking solution (BS: 1% BSA, 0.4% Triton X‐100 in PBS). Following, samples were incubated in primary antibody solutions against H2AX phosphorylated at serine‐139 (γH2AX; 1 : 1000 in BS; Merck Millipore), 53BP1 (1 : 500 in BS; Novus Biologicals), or RAD51 (1 : 250 in BS; Santa Cruz), washed in PBS and incubated in AlexaFluor 555‐conjugated anti‐rabbit (1 : 500 in BS; Abcam) or AlexaFluor 647‐conjugated anti‐mouse (1 : 500 in BS; Thermo Fisher) secondary antibody solutions. Afterward, samples were stained with 1 µg·mL−1 DAPI (Sigma‐Aldrich) and mounted with ProLong Gold Antifade Mountant (Thermo Scientific). Images were examined using a Zeiss AxioImager ApoTome microscope (structured illumination) (Carl Zeiss). zen 2.0 lite software (Carl Zeiss) was used for image analysis.
Alexafluor 647 conjugated anti mouse
Manufactured by Thermo Fisher Scientific
AlexaFluor 647-conjugated anti-mouse is a fluorescent-labeled antibody used in various biological applications. It is designed to detect and visualize mouse-specific targets in samples.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions)
γh2ax, by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Carl Roth (1 mentions)
Axioimager apotome microscope, by Zeiss (1 mentions)
Zen 2.0 lite software, by Zeiss (1 mentions)
Alexafluor 555 conjugated anti rabbit, by Abcam (1 mentions)
Rad51, by Santa Cruz Biotechnology (1 mentions)
Alexa fluor 488 conjugated anti rabbit, by Thermo Fisher Scientific (1 mentions)
Ab5603, by Merck Group (1 mentions)
Ab157524, by Abcam (1 mentions)
Ab6046, by Abcam (1 mentions)
Ab76541, by Abcam (1 mentions)
Hfab rhodamine anti tubulin primary antibody, by Bio-Rad (1 mentions)
2 protocols using alexafluor 647 conjugated anti mouse
1
Immunofluorescence Analysis of DNA Damage Markers
2
Embryonic Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Antibodies used for analysis of embryos include rabbit anti–β-tubulin (Abcam, ab6046, polyclonal, lot: GR31927-5), mouse anti–DICER-1 (Santa Cruz Biotechnology, sc-136981, A-2, lot: I1817), mouse anti–CDX-2 (Abcam, ab157524, CDX2-88, lot: GR300552-6), anti-rabbit anti-CDX2 antibody (Abcam, ab76541, lot #GR300552-18), rabbit anti-SOX-2 (EMD Millipore, AB5603, polyclonal, lot: NG1863962 and 3254559), goat anti–glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Sigma-Aldrich, SAB2500450, polyclonal, lot: 6377C2), rabbit anti-GADD45a (Invitrogen, MA5-17014, D.81.E, lot: R12274975), Alexa Fluor 647 anti-mouse/human CD324 (E-cadherin) antibody (BioLegend catalog no. 147307, lot no. B254794), Alexa Fluor 594 FGFR-1 antibody (Santa Cruz Biotechnology, catalog no. sc-57132 AF594, lot no. J0218), Rhodamine anti–β-actin immunoglobulin G (Bio-Rad, 12004163, lot: 64219909), hFAB Rhodamine anti-tubulin primary antibody (Bio-Rad, 12004165, lot no. 64210670). Donkey secondary antibodies Alexa Fluor 647–conjugated anti-mouse (A31571, lot #1903516), Alexa Fluor 594–conjugated anti-mouse (A21203, lot #2066086), and Alexa Fluor 488-conjugated anti-rabbit (A21206) were purchased from Thermo Fisher Scientific, CA, USA. All bands migrated to approximate molecular masses stated by manufacturers or previously published studies (table S2).
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