Immunofluorescence labeling of tissue specimens was performed by fixing tissue sections or cells in 4% PFA/PBS and blocking in 1% BSA (or permeated with 0.1% Triton X-100/PBS for 10 min for GORAB staining) prior to incubating with primary antibodies. The following primary antibodies were used: GORAB, 1:500 (Proteintech); AQP5, 1:50 (A4979, MilliporeSigma); SFTPC, 1:250 (ab40879, Abcam); VCAN, 1:200 (ab1032, MilliporeSigma); VIM, 1:200 (ab92547, Abcam); F-actin, 1:50 (ab205, Abcam,); VCL, 1:100 (ab129002, Abcam); SCGB1A1, 1:200 (sc-365992, Santa Cruz); ACTA2, 1:500 (a2547, MilliporeSigma); CDH1, 1:100 (BD-610182, BD biosciences). AlexaFluor-conjugated secondary antibodies (1:250) were from Life Technologies (Thermo Fisher Scientific). Sections were sealed in mounting medium with DAPI (Vector Laboratories, Burlingame, CA). TUNEL staining was performed according to manufacturer’s instructions (DeadEnd Fluoremetric, G3250, Promega, Chicago, IL). Images were acquired by Nikon 80i fitted with a Nikon DS-Qi1Mc camera and processed with Photoshop 5.5 CS (Adobe, San Jose, CA).
Gorab
Manufactured by Proteintech
GORAB is a polyspecific antibody that recognizes the GORA protein. It is a useful tool for researchers studying GORA and its associated biological processes.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (3 mentions)
Ds qi1mc camera, by Nikon (3 mentions)
γ tubulin, by Abcam (2 mentions)
Acetylated α tubulin, by Merck Group (2 mentions)
Krt17, by Abcam (2 mentions)
Lectin helix pomatiaagglutinin hpa, by Thermo Fisher Scientific (2 mentions)
Golgin 97, by Thermo Fisher Scientific (2 mentions)
Krt14, by Fortrea (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (2 mentions)
Deadend fluorometric tunel system, by Promega (2 mentions)
Hybond nitrocellulose, by GE Healthcare (2 mentions)
Hrp conjugated secondaryantibodies, by BD (2 mentions)
Supersignal substrate, by Thermo Fisher Scientific (2 mentions)
Polyvinylidene fluoride (pvdf), by Merck Group (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Ab129002, by Abcam (1 mentions)
Ab40879, by Abcam (1 mentions)
Ab205, by Abcam (1 mentions)
A2547, by Merck Group (1 mentions)
Ab92547, by Abcam (1 mentions)
5 protocols using gorab
1
Immunofluorescence Labeling of Tissue Specimens
2
Immunofluorescence Labeling of Tissue Specimens
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Immunofluorescence labeling of tissue specimens was performed as
described previously (Dai et al., 2013 (link)).
Cells were fixed in 4% PFA and blocked in 1% BSA prior to
incubating with primary antibodies. The following primary antibodies were used:
GORAB, 1:500 (Proteintech); KRT14, 1:1,000 (Covance); KRT1, 1:2000 (Roop et al.,
1987); KRT17, 1:200 (Abcam); LOR, 1:500 (Covance); NGFR, 1:200 (Promega); LEF1,
1:100 (Cell Signaling, Danvers, MA); acetylated α-tubulin, 1:600
(Sigma); γ-tubulin, 1:500 (Abcam); ARL13B, 1:100
(#73–287, NeuroMab); lectin Helix pomatiaagglutinin (HPA), 1:1,000 (Invitrogen); Golgin97, 1:1,000 (Molecular Probes),
and Ki67, 1:1000 (BD Pharmingen). AlexaFluor-conjugated secondary antibodies
(1:250) were from Life Technologies. Sections were sealed in mounting medium
with DAPI (Vector Laboratories). TUNEL staining were performed with DeadEnd
Fluorometric TUNEL System (Promega). Images were acquired by Nikon
80i fitted with Nikon DS-Qi1Mc camera and processed with
Photoshop 5.5 CS.
described previously (Dai et al., 2013 (link)).
Cells were fixed in 4% PFA and blocked in 1% BSA prior to
incubating with primary antibodies. The following primary antibodies were used:
GORAB, 1:500 (Proteintech); KRT14, 1:1,000 (Covance); KRT1, 1:2000 (Roop et al.,
1987); KRT17, 1:200 (Abcam); LOR, 1:500 (Covance); NGFR, 1:200 (Promega); LEF1,
1:100 (Cell Signaling, Danvers, MA); acetylated α-tubulin, 1:600
(Sigma); γ-tubulin, 1:500 (Abcam); ARL13B, 1:100
(#73–287, NeuroMab); lectin Helix pomatiaagglutinin (HPA), 1:1,000 (Invitrogen); Golgin97, 1:1,000 (Molecular Probes),
and Ki67, 1:1000 (BD Pharmingen). AlexaFluor-conjugated secondary antibodies
(1:250) were from Life Technologies. Sections were sealed in mounting medium
with DAPI (Vector Laboratories). TUNEL staining were performed with DeadEnd
Fluorometric TUNEL System (Promega). Images were acquired by Nikon
80i fitted with Nikon DS-Qi1Mc camera and processed with
Photoshop 5.5 CS.
3
Western Blot Quantification of Protein Targets
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Protein was extracted in cold RIPA lysis buffer (50 mM Tris-HCl pH 7.4,
150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, and
0.1% SDS) supplemented with proteinase inhibitors. Tissue or cell
lysates were cleared and separated by 10% SDS-PAGE and transferred to
polyvinylidene fluoride (PVDF, Millipore) or Hybond nitrocellulose (GE
Healthcare) membranes, following standard procedures. Blots were probed with the
primary antibodies which were then detected with HRP-conjugated secondary
antibodies (BD biosciences) and SuperSignal substrates (Thermo Scientific).
Enhanced chemiluminescent (ECL) substrate (Pierce, Rockford, IL USA) and
CL-XPosure film (Thermo Scientific) were used for detection. The following
primary antibodies were used: GORAB, 1:1,000 (Proteintech); β-actin,
1:1,000 (Santa Cruz); GLI1, 1:250 (clone V812, Cell Signaling); GLI3 (1
µg/ml, Cell Signaling). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)
or β-actin was used as a loading control. Quantification was performed
with densitometry and ImageJ software (1.43u, NIH).
150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, and
0.1% SDS) supplemented with proteinase inhibitors. Tissue or cell
lysates were cleared and separated by 10% SDS-PAGE and transferred to
polyvinylidene fluoride (PVDF, Millipore) or Hybond nitrocellulose (GE
Healthcare) membranes, following standard procedures. Blots were probed with the
primary antibodies which were then detected with HRP-conjugated secondary
antibodies (BD biosciences) and SuperSignal substrates (Thermo Scientific).
Enhanced chemiluminescent (ECL) substrate (Pierce, Rockford, IL USA) and
CL-XPosure film (Thermo Scientific) were used for detection. The following
primary antibodies were used: GORAB, 1:1,000 (Proteintech); β-actin,
1:1,000 (Santa Cruz); GLI1, 1:250 (clone V812, Cell Signaling); GLI3 (1
µg/ml, Cell Signaling). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)
or β-actin was used as a loading control. Quantification was performed
with densitometry and ImageJ software (1.43u, NIH).
4
Immunofluorescence Labeling of Tissue Specimens
5
Western Blot Quantification of Protein Targets
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