Rabbit anti olfm4, by Cell Signaling Technology - Product details

1

Multicolor Immunostaining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies used in this study are as follows: rabbit anti-Lyz (1:800; Dako, #A0099), rabbit anti-Olfm4 (1:500; Cell Signaling Technology, #39141), recombinant rabbit anti-aldolase B + aldolase C (1:300; Abcam, #ab75751), mouse anti-human KRT20 (1:500; Dako, #M701929-2), mouse anti–Chr-A (1:50; Santa Cruz Biotechnology, #sc-393941), rat anti–E-cadherin (1:400; Santa Cruz Biotechnology, #sc-59778), CD24 Monoclonal Antibody (1:200; Thermo Fisher Scientific, #17-0242-80), and mouse anti-Ki67 (1:200; BD Biosciences, 550609). The secondary antibodies used in this study are as follows: Goat Anti-Rabbit IgG H&L (Alexa Fluor 405) preadsorbed (1:1000; Abcam, #ab175654), Goat Anti-Rat IgG H&L (Alexa Fluor 555) preadsorbed (1:1000; Abcam, #ab150166), Donkey Anti-Mouse IgG H&L (Alexa Fluor 647) (1:500; Thermo Fisher Scientific, #A31571), and Donkey Anti-Rabbit IgG H&L (Alexa Fluor 405) preadsorbed (1:1000; Abcam, #ab175649). The dyes used in this study are as follows: WGA conjugated to CF488A (5 μg/ml; Biotium), RedDot1 Far-Red Nuclear stain (1:200; Biotium), and SYTOX Orange Nucleic Acid Stain (1:5000; Thermo Fisher Scientific, #S11368). The order of staining, optimized to ensure good staining quality for all cell types, was based on the staining quality and stripping difficulty of each antibody (fig. S1G).

Zheng X., Betjes M.A., Ender P., Goos Y.J., Huelsz-Prince G., Clevers H., van Zon J.S, & Tans S.J. (2023). Organoid cell fate dynamics in space and time. Science Advances, 9(33), eadd6480.

+ Open protocol

+ Expand

2

Immunofluorescence Staining of Cell Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Secondary antibodies used are goat anti-rabbit or mouse AlexaFluor 488 or 594, donkey anti-goat or rabbit AlexaFluor 488 or 594 (all from Invitrogen). Sections were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (Sigma, D8417). Primary antibodies used are rabbit anti-active CASPASE 3 (Cell Signaling, 9661), mouse anti-KI67 (BD Pharmingen, 550609), rabbit anti-OLFM4 (Cell Signaling, 39141), goat anti-PTBP1 (Santa Cruz sc-16547), rabbit anti-PTBP1 (gift from Dr. Douglas Black), goat anti-GFP (Rockland, 600-101-215), rabbit anti-P53 (Leica Biosystems, NCL-L-p53-CM5p), rabbit anti-PHLDA3 (LifeSpan BioSciences, LS-C499531-100), rabbit anti-Lysozyme antibody (Diagnostic Biosystems, RP028), and Affinipure Fab Fragment Goat anti-Rabbit IgG (H + L) (Jackson ImmunoResearch, 111-007-003).

Chembazhi U.V., Tung W.S., Hwang H., Wang Y., Lalwani A., Nguyen K.L., Bangru S., Yee D., Chin K., Yang J., Kalsotra A, & Mei W. (2023). PTBP1 controls intestinal epithelial regeneration through post-transcriptional regulation of gene expression. Nucleic Acids Research, 51(5), 2397-2414.

+ Open protocol

+ Expand

3

Multicolor Immunostaining of Tissue Slices

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies used and their respective dilutions were: rabbit anti Olfm4 1:200 (Cell Signaling Technology Cat# 39141), mouse anti CK20 1:50 (Dako Cat# M7019), mouse anti Ki67 1:100 (BD Biosciences Cat# 550609), rabbit anti Lysozyme 1:2000 (Dako Cat# A0099), rabbit anti ZO-1 1:200 (Thermo Fisher Scientific Cat #40-2200) and mouse anti GFP 1:400 (Abcam Cat# ab1218).
The secondary antibodies used were: goat anti mouse Alexa Fluor 488 (Thermo Fisher Scientific Cat# A-11029), donkey anti rabbit Alexa Fluor 488 (Thermo Fisher Scientific Cat# A-21206), goat anti rabbit Alexa Fluor 555 (Thermo Fisher Scientific Cat# A-21429), and goat anti mouse Alexa Fluor 405 (Abcam Cat# ab175660). All secondary antibodies were used at 1:400 dilution. To label F-actin, Phalloidin Atto 488 (Sigma-Aldrich Cat# 49409) was used at 1:500 and Phalloidin Alexa Fluor-647 (Thermo Fisher Scientific, Cat# A22287) was used at 1:400.
For immunostainings of tissue slices, rat anti-E-Cadherin (ECCD-2; Thermo Fisher Scientific Cat# 13-1900) was used at 1:100, DAPI (Sigma-Aldrich Cat# D9542) was used at 5 μg/mL and Rhodamine-phalloidin (Thermo Fisher Scientific Cat# R415) was used at 1:200.

Pérez-González C., Ceada G., Greco F., Matejčić M., Gómez-González M., Castro N., Menendez A., Kale S., Krndija D., Clark A.G., Gannavarapu V.R., Álvarez-Varela A., Roca-Cusachs P., Batlle E., Vignjevic D.M., Arroyo M, & Trepat X. (2021). Mechanical compartmentalization of the intestinal organoid enables crypt folding and collective cell migration. Nature cell biology, 23(7), 745-757.

+ Open protocol

+ Expand

4

Reagents and Materials for Cell Studies

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rabbit anti-LGR4 was purchased from Abcam (ab75501, Cambridge, MA). Rabbit anti-CD36 (18836-1-AP) was purchased from Proteintech (Chicago, USA). Rabbit anti-GLUT2 (A9843), rabbit anti- Presenilin 1(A2187), rabbit anti-LYZ (A13511) and rabbit anti-HES1 (A0925) were from ABclonal (Wuhan, China). Rabbit anti-FATP4 was obtained from Abmart (T57249, Shanghai, China). Rabbit anti-β-catenin was from Cell Signaling Technology (8480, Danvers, MA, USA). Rabbit anti-LaminB1 (12987-1-AP) and mouse anti-β-actin (6009-1-Ig) were purchased from Proteintech (Chicago, USA). Rabbit anti-Ki67 was purchased from Abcam (ab15580, Cambridge, United Kingdom). Rabbit anti-OLFM4 was obtained from Cell Signaling Technology (39141, Danvers, MA, USA). Lgr4 siRNA (5’-GGACUUAUCUUAUAACGAUTT-3’) was synthesized by Synbio Technologies (Suzhou, China). RNAi in vitro transfection reagent was obtained from D-Nano Therapeutics (DN001-10, Beijing, China). TUNEL Apoptosis Assay Kit was purchased from Beyotime Biotechnology (Beyotime, Shanghai, China).

Liang Y., Luo C., Sun L., Feng T., Yin W., Zhang Y., Mulholland M.W., Zhang W, & Yin Y. (2024). Reduction of specific enterocytes from loss of intestinal LGR4 improves lipid metabolism in mice. Nature Communications, 15, 4393.

+ Open protocol

+ Expand

5

Multicolor Immunostaining Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies used in this study are as follows: rabbit anti-Lyz (1:800; Dako, #A0099), rabbit anti-Olfm4 (1:500; Cell Signaling Technology, #39141), recombinant rabbit anti-aldolase B + aldolase C (1:300; Abcam, #ab75751), mouse anti-human KRT20 (1:500; Dako, #M701929-2), mouse anti–Chr-A (1:50; Santa Cruz Biotechnology, #sc-393941), rat anti–E-cadherin (1:400; Santa Cruz Biotechnology, #sc-59778), CD24 Monoclonal Antibody (1:200; Thermo Fisher Scientific, #17-0242-80), and mouse anti-Ki67 (1:200; BD Biosciences, 550609). The secondary antibodies used in this study are as follows: Goat Anti-Rabbit IgG H&L (Alexa Fluor 405) preadsorbed (1:1000; Abcam, #ab175654), Goat Anti-Rat IgG H&L (Alexa Fluor 555) preadsorbed (1:1000; Abcam, #ab150166), Donkey Anti-Mouse IgG H&L (Alexa Fluor 647) (1:500; Thermo Fisher Scientific, #A31571), and Donkey Anti-Rabbit IgG H&L (Alexa Fluor 405) preadsorbed (1:1000; Abcam, #ab175649). The dyes used in this study are as follows: WGA conjugated to CF488A (5 μg/ml; Biotium), RedDot1 Far-Red Nuclear stain (1:200; Biotium), and SYTOX Orange Nucleic Acid Stain (1:5000; Thermo Fisher Scientific, #S11368). The order of staining, optimized to ensure good staining quality for all cell types, was based on the staining quality and stripping difficulty of each antibody (fig. S1G).

Zheng X., Betjes M.A., Ender P., Goos Y.J., Huelsz-Prince G., Clevers H., van Zon J.S, & Tans S.J. (2023). Organoid cell fate dynamics in space and time. Science Advances, 9(33), eadd6480.

+ Open protocol

+ Expand

6

Multicolor Immunostaining of Tissue Slices

Check if the same lab product or an alternative is used in the 5 most similar protocols

The primary antibodies used and their respective dilutions were: rabbit anti Olfm4 1:200 (Cell Signaling Technology Cat# 39141), mouse anti CK20 1:50 (Dako Cat# M7019), mouse anti Ki67 1:100 (BD Biosciences Cat# 550609), rabbit anti Lysozyme 1:2000 (Dako Cat# A0099), rabbit anti ZO-1 1:200 (Thermo Fisher Scientific Cat #40-2200) and mouse anti GFP 1:400 (Abcam Cat# ab1218).
The secondary antibodies used were: goat anti mouse Alexa Fluor 488 (Thermo Fisher Scientific Cat# A-11029), donkey anti rabbit Alexa Fluor 488 (Thermo Fisher Scientific Cat# A-21206), goat anti rabbit Alexa Fluor 555 (Thermo Fisher Scientific Cat# A-21429), and goat anti mouse Alexa Fluor 405 (Abcam Cat# ab175660). All secondary antibodies were used at 1:400 dilution. To label F-actin, Phalloidin Atto 488 (Sigma-Aldrich Cat# 49409) was used at 1:500 and Phalloidin Alexa Fluor-647 (Thermo Fisher Scientific, Cat# A22287) was used at 1:400.
For immunostainings of tissue slices, rat anti-E-Cadherin (ECCD-2; Thermo Fisher Scientific Cat# 13-1900) was used at 1:100, DAPI (Sigma-Aldrich Cat# D9542) was used at 5 μg/mL and Rhodamine-phalloidin (Thermo Fisher Scientific Cat# R415) was used at 1:200.

Pérez-González C., Ceada G., Greco F., Matejčić M., Gómez-González M., Castro N., Menendez A., Kale S., Krndija D., Clark A.G., Gannavarapu V.R., Álvarez-Varela A., Roca-Cusachs P., Batlle E., Vignjevic D.M., Arroyo M, & Trepat X. (2021). Mechanical compartmentalization of the intestinal organoid enables crypt folding and collective cell migration. Nature cell biology, 23(7), 745-757.

+ Open protocol

+ Expand

Rabbit anti olfm4

Lab products found in correlation

Spelling variants of this product

6 protocols using rabbit anti olfm4

Multicolor Immunostaining Protocol

Immunofluorescence Staining of Cell Markers

Multicolor Immunostaining of Tissue Slices

Reagents and Materials for Cell Studies

Multicolor Immunostaining Protocol

Multicolor Immunostaining of Tissue Slices

About PubCompare

Ready to get started?