For imaging flow cytometry experiments: anti-CD8 APC-Cy7 (BD Biosciences, San Jose, CA or Tonbo Biosciences, San Diego, CA) and anti-CD56 PE-CF594 (BD Biosciences) were used for surface staining; all intracellular stains included anti-perforin D48 PE or BV421 (Biolegend, San Diego, CA) and anti-perforin δG9 FITC (Biolegend) and one of the following unconjugated antibodies: anti-CD71 (Cell Signaling Technology, Danvers, MA), anti-granzyme B (BD Biosciences), anti-rab3D (Abcam, Cambridge, MA), anti-rab4 (Pierce Antibodies, Waltham, MA), anti-rab5 (Cell Signaling Technology), anti-rab7 (Santa Cruz Biotechnology, Dallas, TX), anti-rab8 (Cell Signaling Technology), anti-rab11a (Cell Signaling Technology), anti-rab27a (Abcam), anti-rab35 (Abcam), anti-rab37 (Abcam), anti-syntaxin6 (Cell Signaling Technology), anti-syntaxin7 (R&D systems, Minneapolis, MN), anti-vti1b (Abcam), anti-VAMP3 (Abcam), anti-VAMP4 (Abcam), or anti-SNAP23 (Abcam). The unconjugated antibodies were detected using goat anti-rabbit AF647, chicken anti-goat AF647, or donkey anti-sheep AF647 secondary antibodies (Invitrogen, Carlsbad, CA). For confocal microscopy, perforin D48 FITC (Abcam), perforin δG9 AF647 (Biolegend), and goat anti-rabbit or donkey anti-sheep AF568 secondary antibodies (Invitrogen) were used, along with the selected rab or SNARE antibody.
Anti rab27a
Manufactured by Abcam
Sourced in United Kingdom, United States
Anti-RAB27A is an antibody that recognizes the RAB27A protein, a member of the Ras-related protein family involved in vesicle trafficking and secretion. The antibody can be used for the detection and study of RAB27A in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti rab27b, by Abcam (2 mentions)
Anti cd8 apc cy7, by BD (1 mentions)
Anti cd71, by Cell Signaling Technology (1 mentions)
Anti cd56 pe cf594, by BD (1 mentions)
Anti rab5, by Cell Signaling Technology (1 mentions)
Anti rab8, by Cell Signaling Technology (1 mentions)
Anti syntaxin6, by Cell Signaling Technology (1 mentions)
Anti vamp3, by Abcam (1 mentions)
Anti snap23, by Abcam (1 mentions)
Goat anti rabbit af647, by Thermo Fisher Scientific (1 mentions)
Chicken anti goat af647, by Thermo Fisher Scientific (1 mentions)
Anti rab7, by Santa Cruz Biotechnology (1 mentions)
Anti granzyme b, by BD (1 mentions)
Anti rab11a, by Cell Signaling Technology (1 mentions)
Anti rab37, by Abcam (1 mentions)
Phalloidin 568, by Thermo Fisher Scientific (1 mentions)
Prolong diamond antifade reagent, by Thermo Fisher Scientific (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Skim milk, by Merck Group (1 mentions)
Anti alix antibody, by Abcam (1 mentions)
4 protocols using anti rab27a
1
Imaging Flow Cytometry for Immune Cell Analysis
2
Cytotoxic T Cell Degranulation Assay
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CTLs were washed and allowed to adhere for 12 minutes onto hydrophobic mutliwell slides (Hendles-Essex) in FCS-free IMDM. For conjugation assays, blue P815 target cells were coated with anti-CD3 (UCHT1), plated on multiwall slides in serum free media and CTLs were added for 10-15min. Cells were fixed with 4% paraformaldehyde (15710-S, Electron Microscopy Systems, USA) and incubated in quenching solution (50 mM ammonium chloride in 1X PBS) (15 min at RT) and rinsed twice in PBS. Permeabilization was done with 0.2% Saponin in PBS (5 min, RT) followed by washing and blocking for 30min in blocking buffer (1xPB, 1%BSA, 0.2% saponin). Cells were stained with anti-LAMP1 (H4A3, hybridoma supernatant), anti-RAB27A (abcam, UK) for 1 hour, followed by fluorophore-conjugated secondary antibodies (donkey anti-mouse 488 and goat anti-rabbit 647, Invitrogen, UK) (1 hour at RT) together with phalloidin 568 (Invitrogen, UK). Nuclei were stained for 5 minutes at RT with Hoechst (Thermo Fisher, UK), and samples were mounted in ProLong Diamond Antifade Reagent (Thermo Fisher, UK).
3
Western Blot Analysis of Exosomal Markers
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After quantitative analysis, the total protein of the sample was extracted and denatured in this study. SDS–PAGE gel was used for electrophoresis, electrophoresis apparatus (Bio‐RAD) was adjusted to 120 V for electrophoresis, PVDF membrane (Millipore) was used for membrane transfer, and skim milk (Sigma) was used for blocking. Primary antibodies (Abcam) were then added and incubated overnight. The antibodies used were as follows: anti‐Rab11 (Abcam, 1:1000), anti‐Rab27a (Abcam, 1:200), anti‐Rab27b (Abcam, l:500), anti‐TSAP6 antibody (Abcam, 1:1000), anti‐CD63 antibody (Abcam, 1:1000), anti‐CD9 antibody (Abcam, 1:1000), anti‐Alix antibody (Abcam, 1:1000), and anti‐β‐Actin antibody (Abcam, 1:10,000). The membrane was then incubated with the primary antibody diluted in advance. Subsequently, the membranes were incubated with IgG (Abcam, 1:300) for 1 h. Finally, the protein bands were visualized using a Pierce ECL Western blotting kit. Protein expression was quantified using ImageJ to analyze band gray values. β‐Actin was used as a control.
4
Comprehensive Protein Analysis in Cellular Signaling
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Protein levels were analyzed using IB and ELISA following conventional protocols. For IB, the primary antibodies used were anti-YAP (Abcam, Cambridge, MA, USA, #ab52771), anti-GAPDH (CST, Boston, MA, USA, #5174 and #51332), anti-p-127-YAP (Abcam, #ab76252), anti-p-381-YAP (CST, #13619), anti-O-Glc-YAP (developed by Biolynx, Hangzhou, China), anti-HRS (Abcam, #ab72053), anti-PLD2 (CST, #13904), anti-RAB2B (Novus, Littleton, CO, USA, #NBP1-31631), anti-RAB27A (Abcam, #ab55667), anti-RAB27B (Abcam, #ab103418), anti-VAMP (Abcam, #ab36195), anti-ATG7 (Abcam, #ab52472), anti-SOX10 (Abcam, #ab227680), anti-Mycn (Abcam, #ab227822), anti-ALYREF (Abcam, #ab202894), anti-NSUN2 (Abcam, #ab259941), anti-DNMT2 (Abcam, #ab220175), anti-NSUN1 (Abcam, #ab270175), anti-NSUN3 (Abcam, #ab272616), anti-NSUN4 (Abcam, #ab235430), anti-NSUN5 (Abcam, #ab121633), anti-NSUN6 (Novus, #NBP1-92202), anti-NSUN7 (Biorbyt, Cambridge, UK, #orb258175), anti-YBX1 (Abcam, #ab255606), anti-EXOSC10 (Abcam, #ab94981), anti-SPI1 (Abcam, #ab227835), anti-CD63 (Abcam, #ab271286), anti-TSG101 (Abcam, #12501), anti-ALIX (Abcam, #ab275377), anti-CD9 (Abcam, #ab92726), and anti-calnexin (Abcam, #ab133615). For ELISA, YAP and NSUN2 levels were measured using kits from Yingxin Biotech Ltd. (Shanghai, China) as per manufacturer’s instructions.
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